PBG 201 Principles of Genetics and Cytogenetics (2+1)

Theory Notes

Lec 1: Definition of genetics, heredity, inheritance, cytology, cytogenetics; Brief history

of developments in genetics and cytogenetics
Genetics : A biological science which deals with the study of heredity (inheritance) and variation.
It is the study of structure, composition, transmission and function of genes. The term genetics was
coined by [Link] (1905).
Heredity / inheritance: Transmission of characters from parents to their offspring
Variation: Differences among the individuals of a species for a character (trait). It may be
(i) Heritable (genetic) and (ii) Non-heritable (environmental)
Heritable variation: Differences in inherited traits transmitted from generation to generation
Environmental variation: Temporary and not inherited to next generation
Cytology: Branch of biology deals with the structure and function of cells.
Cytogenetics: Branch of genetics deals with the study of structure, function and behavior of
chromosomes in relation to gene inheritance.
Mendelian genetics: Genetics which deals with the inheritance of oligogenic characters or
qualitative characters which displays discontinuous variation.
Non-Mendelian genetics: It involves the study of role of cytoplasm and its organelles in heredity.
Quantitative genetics: Genetics which deals with the inheritance of quantitative or polygenic
characters which displays discontinuous variation.
Mutation genetics: It includes the study of chromosomal changes and gene mutations.

History of genetics and cytogenetics

Pre Mendelian period

 The [Link] (1761) performed first hybridization in tobacco. He demonstrated that the
hybrids resemble any one of the other parent or intermediate between them. Both the
parents make equal contributions to the hybrids
 Knight (1779) conducted experiments on pea much before Mendel but failed to
formulate the laws of inheritance because he could not use the mathematics
 Gartner (1772-1850) and Naudin (1815-1899) done experiments similar to Kolreuter
and they observed the similar results.
Mendelian period
History of genetics started with the work of the Austrian monk Gregor Johann Mendel. He
studied botany in the University of Vienna and Worked with pure lines of garden pea for eight
years. Prior to Mendel, heredity was regarded as a "blending" process and the offspring were
essentially a "dilution "of the different parental characteristics.
He laid the foundation of the science of genetics in 1866 and discovered the basic
principles of heredity. He has formulated two important laws of inheritance, viz., (i) law of
segregation and (ii) law of independent assortment
 His work on pea plants, published in 1866, as Mendelian inheritance. Mendel has
presented two papers in “Natural History Society” in 1865 (Published in proceedings of the
society in 1866) and major findings are
1. Inheritance of each trait is determined by "factors" that are passed on to descendents
unchanged
2. Individual inherits one such factor from each parent for each trait
3. A trait may not show up in an individual but can pass on to the next generation
The year 1900 marked the "rediscovery of Mendel" by Hugo de Vries, Carl Correns and
Erich von Tschermak.

Post-Mendelian period
Fredrich Miescher (1869) has successfully isolated nuclein from pus cells obtained from
discarded [Link] noticed that nuclein was acidic, contained a lot of phophorus and
nitrogen and was found in the nucleus of cells. Nuclein later on known as DNA.
Swiss botanist Nageli (1840) first described thread-like structures in the nuclei of plant cells,
and he called as “transitory cytoblasts” are now known as chromosomes.
Later, Waldeyer (1888) coined the term “chromosome” after staining techniques had been
developed to make them more visible (chromos = colour; soma = body).
Theodor Boveri (1888-1890) has investigated the chromosomes and explained that
chromosomes are organized individual structures throughout the process of cell division. He also
stated that sperm and egg contribute the same number of chromosomes.
Walter Sutton (1902 ) along with Boveri, proposed Sutton- Boveri Chromosome Theory. He
suggested that chromosomes are paired and may be the carriers of heredity and Mendel’s
"factors" are located on chromosomes.
Wilhelm Johannsen (1909) has introduced the terms Gene, Genotype and Phenotype.

The basic principles of Mendelian genetics had been applied to a wide variety of
organisms, most notably the fruit fly Drosophila melanogaster during 1915 by Thomas Hunt
Morgan. Morgan proved that genes are carried on chromosomes. He also demonstrated the
existence of sex-linked genes and established the chromosome theory of heredity.
Fred Griffith (1928) has studied different strains of pneumococci, the bacteria that can
cause pneumonia. This bacteria present in two strains viz., S and R. S-form has a capsule and
looks smooth under the microscope. It is virulent and kills infected mice because the immune
system cannot break through the cell wall of the S bacterium.
Phoebus Levene (1929) has identified the DNA deoxyribose sugar in “thymus” nucleic
acid and the RNA ribose sugar in “yeast”nucleic acid. He has also identified the nitrogenous
bases: adenine, guanine, cytosine and thymine in “thymus” nucleic acid and uracil in “yeast”
nucleic acid.
Avery, MacLeod and McCarty (1944) has explained the significance of DNA and proved
that DNA is the genetic material
Erwin Chargaff (1950) discovered that in DNA, the amount of Adenine is equal to the
amount of Thymine and the amount of Guanine is equal to the amount of Cytosine.
Chargaff”s Rule: A=T & G=C
 Rosalind Franklin (1951-1953) was responsible for much of the research and discovery
work that led to the understanding the structure of deoxyribonucleic acid, DNA. She is the brilliant
chemist whose X ray diffraction studies provided crucial clues to the structure of DNA.
James Watson and Francis Crick (1953) approach was to make physical models to
narrow down the possibilities and eventually create an accurate picture of the molecule. They
suggested that the DNA molecule was made of two chains of nucleotides, each in a helix (as
Franklin had found) but one going up and the other going down. Specifically, the large adenine
molecule could pair with only the smaller thymine and the large guanine molecule could pair
with only the smaller cytosine.
Arthur Kornberg (1957) discovered and isolated DNA polymerase, which becomes the
first enzyme used to make DNA in a test tube. He also proves that the strands are anti-parallel
and that replication proceeds only in one direction (5’to3’).
Meselson and Stahl (1958) experiment determined the mechanism of DNA replication.
Nathan & Smith (1970) isolated first restriction endonuclease enzyme that cuts DNA molecules
at specific sites. This allows scientists to create clones and observe their function.
Normen Borlaug (1975) involved in the development of dwarf wheat
Jeffreys and Flavell (1977) provided the concept of introns and exons.
Nirenberg & Khorana (1978) has made complete genetic code
Muller & Smith (1983) - creation of polymerase chain reaction (PCR) through a combination of
the computer revolution and more easily available enzymes. These in turn would make the job
of mapping the entire human genome possible.
In the 1940s and early 1950s, experiments proved that DNA as the portion of
chromosomes which held genes. A focus on new model organisms such as viruses and bacteria,
along with the discovery of the double helical structure of DNA in 1953, marked the transition to
the era of molecular genetics.
In the following years, chemists developed techniques for sequencing both nucleic acids
and proteins, while others worked out the relationship between the two forms of biological
molecules: the genetic code. The regulation of gene expression became a central issue in the
1960s; by the 1970s gene expression could be controlled and manipulated through genetic
engineering. In the last decades of the 20th century, many biologists focused on large-scale
genetics projects, sequencing entire genomes.
Lec 2: Physical basis of heredity: Structure and function of cell and cell organelles

Definitions
Cell: A basic unit of structure and function in all living organisms.
Cell organelles: Various membrane bound structures that are found within a cell.
Nucleus: In eukaryotes, a double membrane, oval or spherical structure which contains
chromosomes. It regulates growth and reproduction of the cell.
Cell membrane / plasma membrane: Differentially permeable membrane through which extra
cellular substances may be selectively sampled and cell produces may be liberated.
Nuclear membrane: A double membrane outer boundary of the nucleus. Provides selective
continuity between nuclear and cytoplasmaic materials.
Nucleolus: A spherical body found within the nucleus
Chromatin: Partly clumped and tangled mass of nuclear chromosomes
Plastids: self replicating cytoplasmic organelles found in the plant cells.
Leucoplasts: Colourless plastids which are associated with storage of starch, protein and fat.
Chromoplasts: Plastids with other than green colour.
Chloroplast: Plastids of green colour that area associated with photosynthesis.
Grana : Small cylindrical structures found inside the inner membrane of a chloroplast.
Stroma: The space found inside the inner membrane of a chloroplast.
Mitochondria: A rod like cytoplasmic organelle which is the main site of cellular respiration.
Endoplasmic reticulum: A vast network of membranes enclosed tubules, vesicles and sacs found
in the cytoplasm.
Ribosomes: Small cellular particles that are the sites of protein synthesis.
Lysosomes: Cellular particles which contain several digestive enzymes.
Cell wall: The outer most part of a plant cell. Thick cellulose wall surrounding the cell membrane
gives strength and rigidity to the cell.
Middle lamella: A common layer found between adjacent cells.
Primary cell wall: A thin and elastic membrane which lies between middle lamella and secondary
cell wall.
Secondary cell wall: the innermost layer of cell wall which lies between primary cell wall adn
plasma membrane.
Golgi bodies: A cell organelle which is associated with the packaging of food materials such as
proteins, lipids and phospholipids for transport to other cells.
Vacuoles: Storage depots for excess water, waste products, soluble pigments, etc
Tonoplast: A vascular membrane surrounding a vacuole.
Cristae: A series of inside folds in mitochondria.
Cytoplasm: The portion of cell other than nucleus
Hyaloplasm: The portion of cytoplasm other than cell organelles.
Cell is the structural and functional unit of all living organisms and building block of life. In 1665,
Robert Hooke, an Englishman examined thin slices of cork and observed that it was composed of
numerous little boxes, fitted together like honey comb. He named these boxes as cells. The cork
cells studied by Hooke were really empty boxes; they had lost their living matter, the protoplasm.
After his discovery, the protoplasm in living cells was largely over looked due to its transparency.
Today, with the help of special techniques, we could able to see not only the protoplasm but also
many organelles inside it.

 The word cell comes from the Latin word cellula

 Organisms are unicellular / multicellular
 Cells may be spherical, cylindrical, rod-shaped, hexagonal or of irregular shape
 Size of cells ranges from 0.2 to 5.0 µ (micrometers (µm)) in bacteria to 75 mm in the case
of ostrich eggs (without shell)
 Typical plant cell size is 10 – 100 μm

The cell theory, developed in 1839 by Matthias Jakob Schleiden and Theodor Schwann, states
that
 All organisms are composed of one or more cells.
 All cells come from preexisting cells.
 Vital functions of an organism occur within cells, and
 All cells contain the hereditary information necessary for regulating cell functions and for
transmitting information to the next generation of cells.

A cell is made up of following components/organelles:

I. Cell Wall
II. Plasma membrane
III. Cytoplasm – consists of following organelles
1. Mitochondria
2. Endoplasmic reticulum
3. Ribosomes
4. Golgi apparatus
5. Plastids (plant cell)
6. Lysosomes
7. Peroxisomes
8. Vacuoles
9. Centrioles (animal cell)
IV. Nucleus

Cell Wall
Plant cells are surrounded by a non living and rigid coat called cell wall. The cell wall is
present only in plant cell. Though the cell wall is not a living part of the cell, it is an extra
cytoplasmic product.
 Plant cell

 Found outside the cell membrane in plant cells

 Provide plant cells a definite shape
 Provides mechanical support and strength to tissues and organs
 Protection to the cell membrane (plasma membrane)
 It is a structural barrier to some molecules and invading insects.

Cell wall is divided into 3 distinct parts:

(1) middle lamella
(2) primary cell wall and
(3) secondary cell wall

1. Middle lamella
i. The walls of contiguous (immediate neighbour) cells are joined by middle lamella.
ii. The middle lamella is rich in pectins occurring most likely in the form of calcium (Ca2+) and
magnesium (Mg2+) salts.
iii. Adhesion of the walls of contiguous cells is primarily dependent on the presence of Ca2+
and Mg2+ ions.
2. Primary cell wall
i. Primary cell wall is deposited after the formation of middle lamella
ii. Thin, flexible layer formed while the cell is growing.
iii. Main constituents are hemicellulose (53%) and cellulose (30%); in addition, it contains
pectin (5%), protein (5%) and lipid (7%)
iv. Several cellulose molecules associate to form a cellulose microfibril
v. Cellulose microfibrils are the units of cell wall structural organisation.
3. Secondary cell wall
i. The secondary cell wall, a thick layer formed inside the primary cell wall after the cell is
fully grown.
ii. Secondary cell wall is the last to be deposited and lies between primary cell wall and
plasma lemma; in a cell it is the innermost layer of wall.
iii. It is composed mainly of cellulose, but in many tissues it contains lignin, suberin and, in
some cases, cutin.
 iv. Several microfibrils associate to form a macrofibril, which is the structural unit of secondary
cell walls.

Plasma Membrane / cell membrane

i. All living cells, prokaryotic and eukaryotic, have a plasma membrane that encloses their
contents and serves as a semi-porous barrier to the outside environment.
ii. The plasma membrane (cell membrane) is made of two layers of phospholipids. The
membrane has many proteins embedded in it.
iii. The plasma membrane regulates what enters and leaves the cell.
iv. The plasma membrane is semi permeable to specific molecules, however, and allows
nutrients and other essential elements to enter the cell and waste materials to leave the cell.
v. Small molecules, such as oxygen, carbon dioxide, and water, are able to pass freely across
the membrane, but the passage of larger molecules, such as amino acids and sugars, is
carefully regulated. Molecules cross the cell membrane by diffusion and osmosis.

Plasmodesmata

 Plasmodesmata are small channels that directly connect the cytoplasm of neighbouring
plant cells to each other, establishing living bridges between cells.
 These small passages penetrate the middle lamella as well as the primary and secondary
cell walls, providing pathways for transporting cytoplasmic molecules from one cell to
another and are important in cellular communication.
  Plasmodesmata are lined with the plasma membrane so all connected cells are united
through essentially one continuous cell membrane.
 A majority of plasmodesmata also contain a narrow tube-like structure called the
desmotubule, which is derived from the smooth endoplasmic reticulum of the connected
cells.

Cytoplasm
 Part of cells outside the nucleus / the substance except nucleus surrounded by the
plasmamembrane known as cytoplasm
 Cytoplasm is often used to refer to the jellylike matter (termed as cytosol) in which the
organelles are embedded.
 Most of the activities in the cytoplasm are chemical reactions (metabolism), for example,
protein synthesis.
In many cells, the cytoplasm is made up of two parts:
i. Ectoplasm (or plasmagel), a dense gelatinous outer layer concerned with cell movement,
ii. The endoplasm (or plasmasol), a more fluid inner part where most of the organelles are
found.

Organelles of the cytoplasm

1. Mitochondria
 Mitochondria (Mitos – thread; chondria – granule) are rod-shaped organelles considered
as the power generators of the cell.
 Mitochondria, which are found in nearly all eukaryotes, including plants, animals, fungi
 Two specialized membranes encircle each mitochondrion, dividing the organelle into a
narrow intermembrane space and a much larger internal matrix, each of which contains
highly specialized proteins.
 The outer membrane of a mitochondrion contains many channels formed by the protein
porin and acts like a sieve, filtering outer molecules.
 The inner membrane, which is highly convoluted so that a large number of infoldings called
cristae are formed, also allows only certain molecules to pass through it and is much more
selective than the outer membrane.
 Functions
i. Mitochondria are the power
generators of the cell, converting
oxygen and nutrients into
adenosine triphosphate (ATP). ATP
is the chemical energy "currency"
of the cell that powers the cell's
metabolic activities.
ii. Sites of cell respiration. Oxidation
of carbohydrates, lipids and
protein occur in mitochondria.

2. The Endoplasmic Reticulum

 Introduced by Poter and Kallman (1952). It is a continuous system connected on one side
to plasma membrane and on the other side to the nuclear envelope
 Endoplasmic means "within the plasm" and reticulum means "network". All eukaryotic cells
have endoplasmic reticulum.
 It is a network of flattened sacs (cisternae) and branching tubules that extends throughout
the cytoplasm in plant and animal cells.
 These sacs and tubules are all interconnected by a single continuous membrane so that the
organelle has only one large, highly convoluted and complexly arranged lumen (internal
space). Usually referred to as the endoplasmic reticulum cisternal space, the lumen of the
organelle often takes up more than 10 percent of the total volume of a cell.

Functions
 It allows molecules to be selectively transferred between the lumen and the cytoplasm,
and since it is connected to the double-layered nuclear envelope, it further provides a
pipeline between the nucleus and the cytoplasm.
 It manufactures, processes, and transports a wide variety of biochemical compounds for
use inside and outside of the cell.
There are two basic kinds of endoplasmic reticulum based on morphology:
 1. Rough ER: The surface is covered with ribosomes. This type of ER is involved mainly in
the production and processing of proteins that will be exported, or secreted, from the
cell.
2. Smooth ER: Involved in the production of lipids (fats), building blocks for carbohydrate
metabolism, and the detoxification of drugs and poisons.

3. Ribosomes
 It is the protein factory of the cell.
 All living cells contain ribosomes, tiny organelles composed of approximately 60 percent
ribosomal RNA (rRNA) and 40 percent protein.
 Ribosomes are mainly found bound to the endoplasmic reticulum and the nuclear envelope,
as well as freely scattered throughout the cytoplasm.
 The eukaryotic cell generally contains 80S ribosomes and is comprised of 40s and 60s
subunits.
 Prokaryotic cells, on the other hand, contain 70S ribosomes, each of which consists of a 30s
and a 50s subunits.

Functions
 Protein synthesis requires the assistance of two other kinds of RNA molecules in addition to
rRNA. Messenger RNA (mRNA) provides the template of instructions from the cellular DNA
for building a specific protein. Transfer RNA (tRNA) brings the protein building blocks,
amino acids, to the ribosome. There are three adjacent tRNA binding sites on a ribosome:
the aminoacyl binding site for a tRNA molecule attached to the next amino acid in the
protein, the peptidyl binding site for the central tRNA molecule containing the growing
peptide chain, and an exit binding site to discharge used tRNA molecules from the
ribosome.
4. Golgi apparatus (The shippers)
 In 1898, an Italian physician Camilio Golgi discovered the Golgi bodies. Also called as
lipochondria or dictyosomes.
 The Golgi apparatus (GA) also called Golgi bodies or Golgi complex.
 It is typically comprised of a series of five to eight cup-shaped, membrane-covered sacs
called cisternae that look like a stack of deflated balloons.
  Animal cells generally contain 10-20 Golgi stacks per cell, which are linked into a single
complex by tubular connections between cisternae. This complex is usually located close to
the cell nucleus.
 Each Golgi stack has two distinct ends, or faces. The cis face of a Golgi stack is the end of
the organelle where substances enter from the endoplasmic reticulum for processing, while
the trans face is where they exit in the form of smaller detached vesicles.

Functions:
i. The Golgi apparatus is often considered as the distribution and shipping department for
the cell's chemical products.
ii. It packages, modifies, and transports proteins and lipids (fats) to different location
inside/outside of the cell.
iii. In addition, it manufactures a variety of macromolecules on its own, including a variety of
polysaccharides.

5. Plastids
Plastids are found in the cytoplasmic matrix of plant cells only. These structures are
generally spherical or ovoid in shape and they are clearly visible in living cells. 3 types of
plastids found in plant cells:
i. Chromoplasts
Chromoplasts are red, yellow or orange in colour and are found in petals of flowers and
in fruit. Their colour is due to two pigments, carotene and xanthophyll.

Functions
The primary function in the cells of flowers is to attract agents of pollination, and in fruit to
attract agents of dispersal.

ii. Leucoplasts
Leucoplasts are colourless plastids and occur in plant cells not exposed to light, such as
roots and seeds. They are colourless due the absent of pigments.
Functions
Leucoplasts are the centers of starch grain formation; they are also involved in the
synthesis of oils and proteins.
iii. Chloroplasts
 The green colour of chloroplasts is caused by the green pigment chlorophyll.
 Chloroplasts are enclosed by two concentric membranes, each being about 50 Å thick.
 The membrane-free space enclosed by these two membranes is referred to as stroma.
Stroma, a semi-fluid material that contains dissolved enzymes and comprises most of the
chloroplast's volume.
 In higher plants, pile of cylindrical structures in stroma is known as grana. Each granum
contains 5-25 thylakoids (closed hollow disks) stacked on top of each other. The numerous
thylakoids in each stack are connected via their lumens (internal spaces).
 Like mitochondria, chloroplasts possess their own chloroplast DNA and special ribosomes
and RNAs as well.

Functions
i. Converting light energy into
chemical energy through
photosynthesis.
ii. The chemical energy that is
produced by chloroplasts is finally
used to make carbohydrates like
starch, that get stored in the plant.

6. Lysosomes (Clean-up Crews)

 Christian de Duve (1949) has discovered lysosomes. Vesicles of 400-800 mµ produced
from the Golgi complex;
 Membrane bound vesicles containing around 40 hydrolytic enzymes necessary for
digesting certain material in a cell. The main enzyme present in lysosomes is acid
phosphatase; other enzymes are acid DNAase, acid RNAase, β-galactosidase, etc.
 Contain an ionic pump which maintains a highly acidic pH.

Two types

Primary lysosomes are produced by Golgi bodies and contain hydrolytic enzymes only.
They fuse with food vacuoles produced through phagocytosis and pinocytosis to generate
secondary lysosomes.
Therefore, secondary lysosomes contain both hydrolytic enzymes as well as food materials.

Functions
i. Digestion of macromolecules which enter
the cell. The lysosomes infuse with vesicles of
engulfed material and release the digestive
enzymes to break up the material.
ii. Involved in cellular digestion. When a cell
dies, lysosomes rupture and releases
enzymes. These enzymes then digest the
dead cell (autodigestion).
7. Peroxisomes
 They called as peroxisomes because they generate and break down hydrogen peroxide.
 It is a self replicating organelle containing oxidative enzymes. They are similar to
lysosomes.
 They are associated with glycolate metabolism in photosynthesis.
Functions
 to convert fats to carbohydrates
 to detoxify potentially harmful molecules which form in the cell.

[Link]
 Plant cells have one or more vacuoles of variable size.
 In mature and differentiated cells, the major part (upto 90%) of cytoplasm is occupied by
a large vacuole, and the cytoplasm is pushed to the periphery of cells.
 The material contained in the vacuoles is referred to as cell sap.
 Surrounded by a unit membrane; this membrane is referred to as tonoplast.

Functions
 Storage of water
 Function to maintain high internal water
pressure, which aids in the physical
support of plant tissues

9. Centrioles
 Centrioles are cylindrical structure of about 1200-1500 Å in diameter and of about
3,000 - 5,000 Å in length.
 They are confined to animal cells, and are not found in plant cells. In plant cells, structure
similar to centrioles are found at the base of flagella.
 These paired organelles are typically located together near the nucleus. The cytoplasm at
the poles of spindle is known as centrospheres. The centrioles and centrospheres together
knowa as centrosomes.
 A pair of centrioles lies at each of the two poles in a cell from which spindle fibres
radiate toward the equatorial plate.
Functions
 Centrioles organize the spindle apparatus on which the chromosomes move during
mitosis.

NUCLEUS: The semifluid matrix found inside the

nucleus is called nucleoplasm. Within the
nucleoplasm, most of the nuclear material consists
of chromatin, the less condensed form of the cell's
DNA that organizes to form chromosomes during
mitosis or cell division. The nucleus also contains
one or more nucleoli, and a variety of other
smaller components, such as Cajal bodies, GEMS
(Gemini of coiled bodies), and interchromatin
granule clusters.
Three distinct structures
Nuclear envelope, Nucleolus and Chromatin fibers
Nuclear envelope
Nucleus is enclosed by two concentric membranes, each being 70-80 Å in thickness. There is a
space of about 200-300 Å between the two membranes (perinuclear space). The two
membranes, together with the perinuclear space, are known as nuclear envelope.
Nucleolus
The nucleolus looks like a large dark spot within the nucleus. A nucleus may contain up to four
nucleoli, but within each species the number of nucleoli is fixed. After a cell divides, a nucleolus is
formed when chromosomes are brought together into nucleolar organizing regions. During cell
division, the nucleolus disappears. It helps in the production and organisation of ribosomes.
Chromatin fibres
Chromatin fibers contain about 40% DNA, 55% protein and 4-5% RNA. They are the basic units
of chromosome structure. They are also the fundamental basis of inheritance since they contain the
genetic material, viz., DNA.
It produces ribosomes, tRNA, mRNA and other RNAs, which produces the various structural and
enzymatic proteins

Nuclear Pores
The nuclear envelope is perforated with holes called nuclear pores. These pores regulate the
passage of molecules between the nucleus and cytoplasm.

Functions:
 It serves as the information processing and administrative centre of the cell.
 It stores the cell's hereditary material i.e. DNA.
 It coordinates the cell's activities, which include growth, intermediary metabolism, protein
synthesis and reproduction (cell division).
Lec 3: Differences between Prokaryotes and Eukaryotes;
Cell division – mitosis and its significance

Prokaryote: Unicellular organisms whose cells lack membrane bound nucleus like bacteria,
bacteriophage and blue green algae.
Plasmid: Small circular DNA strand (genetic information) found in prokaryote
Eukaryote : Organisms whose cells contain well defined nucleus
Cell division: The process of reproduction of new cells from the pre-existing cells.
Mother cell: The cell which undergo division.
Daughter cells: The new cells which are formed by the process of cell division.
Mitosis: The cell division which produces two identical daughter cells from a mother cell.
Cell cycle: The period in which one cycle of cell division is completed. It consists of interphase and
mitotic phase.
Interphase: A stage in which DNA synthesis takes place. It lies between telophase and prophase.
G1 phase: A pre-DNA replication phase. It lies between telophase and S phase.
S Phase: DNA replication phase. It is between G1 and G2 phases.
G2 phase: A post DNA replication phase during which protein and RNA synthesis take place.
Karyokinesis: The process of division of nucleus.
Cytokinesis: the process of division of cytoplasm.
Differences between eukaryotes and prokaryotes

Eukaryotic cells Prokaryotic cells

Eukaryotes are higher organisms Prokaryotes are primitive organisms

(Eu = good or true; Karyon = nucleus) They (Pro = primitive; Karyon = nucleus)
are generally multi-cellular They are generally uni-cellular
Nucleus is enclosed by two concentric unit The region containing chromosome is not
membranes surrounded by a membrane
DNA is complex with histone and non-histone DNA is naked and not complexed with histones
proteins to form chromatin fibres
DNA per cell is many fold greater Small fraction

DNA is linear and lies within the nucleus DNA is circular and lies free in the cytoplasm
Cell division is mitosis and meiosis Cell division is amitosis (binary fission)

Cytoplasm contains : ER, GB, lysosomes No ER and other membranous structures

Contain Mitochondria Absent : oxidative phosphorylation associated

with plasma lemma
Chloroplast which have typical grana Absent; BGA have lamellar photosynthetic
structure
Cytoplasm has microtubules No microtubules
Ribosomes are 80 S size; in plants they 70 S and made up of 50 S and 30 S
dissociate into 60 S and 40 S
Spindle apparatus is organized Absent
Chromosome ends are attached to the Attached to plasma lemma
nuclear membrane
Most ribosomes are attached to ER All ribosomes are free
Nucleolus present Absent

Cell division usually occurs as part of a larger cell cycle. A cell cycle consists of two phases
viz.,interphase and the cell division. The time required for the completion of cell cycle differs from
species to species.

Interphase

G1 phase (pre-DNA replication), S phase (DNA replication) and G2 phase (post-DNA replication).
Activation of each phase is dependent on the proper progression and completion of the previous
one. Cells that have temporarily or reversibly stopped dividing are said to have entered a state
of quiescence called G0 phase.
G1 phase: Pre-DNA replication phase lies between telophase and S phase. Metabolic changes
prepare the cell for division. Longest phase is observed in Vicia faba (12 hours).Most variable
period of cell cycle. Synthesis of proteins and RNA takes place during this phase. Cells increase in
size.
S phase: This phase comes after G1. Chromosome and DNA replication takes place in this phase.
Each chromosome now consists of two sister chromatids.
G2 phase: Gap between DNA synthesis and mitosis. The cell will continue to grow during this
phase. Metabolic changes assemble the cytoplasmic materials necessary for mitosis and
cytokinesis.

Control of the cell cycle

The cell cycle is controlled at three checkpoints to minimize occurrence of mistake in cell cycle
progression.

 During G1 prior to the S stage

 During G2 prior to the M stage
 During the M stage prior to the end of mitosis

Cell division
Cells are produced by the division of pre-existing cells. Division of nucleus is called karyokinesis
and division of cytoplasm is called cytokinesis.

Types of cell division:

1. Mitosis and [Link]
1. Mitosis
Walther Flemming coined the term mitosis in 1882. It occurs in somatic organs like root tip,
stem tip, leaf base also known as somatic cell division. Daughter cell is similar to mother cell in
all aspects. This is known as homotypic or equational division. Mitosis is the mechanism in
which the chromosome content of a somatic cell (haploid or diploid) is kept constant
through successive cell divisions. The division of the cell is initiated by division of the nucleus i.e.
karyokinesis followed by division of cytoplasm i.e. cytokinesis. The stages of karyokinesis
are – prophase, metaphase, anaphase and telophase.
i. Prophase:
Size of the nucleus is comparatively big. In the resting nucleus the chromatin is spread out as
a network. Gradually the chromosome becomes thick and condensed. The chromatids remain
coiled around each other spirally throughout their length. The chromosomes become distinct as
individual units due to coiling. The nucleolus and nuclear membrane gradually disappear.
ii. Metaphase:
A new structure, the spindle fibres appears in the cytoplasm, which chemically, consists of long
chain protein molecules oriented longitudinally between two poles. Centromere of each
chromosomes is connected to the poles through spindle fibres. Chromosomes moves towards
equatorial plate and arranged in equatorial plane known as auto orientation. Chromosomes
are short and thickest.
iii. Anaphase:
The chromosomes split in the centromere and the sister chromatids are pulled by the
spindle fibers to opposite poles of the cell. At the termination of anaphase, chromosomes
form densely packed group at the two poles. This helps in identical division of chromatids and
 the number of chromosomes remains constant but the quantity of each chromosome is reduced
to half.

iv. Telophase:
This is reorganization phase resulting in the formation of two daughter nuclei. Nuclear
membrane and nucleoli reappear and surround the chromosomes. The newly formed nucleus
contains the same numbers of chromosomes, as this was in parent nucleus.

Cytokinesis:
Just after the nuclear division, the division of cytoplasm takes place which is known as cytokinesis.
The cytokinesis takes place in two ways. In plants, cytokinesis takes place through the formation of
cell plate, which begins in the centre of cell and moves towards the periphery in both sides. In
animals, this occurs by a process known as cleavage, forming a cleavage furrow.
in equatorial region that deepens to form a wall separating the two daughter nuclei.
Significance of Mitosis:
i. Genetic stability (purity): Produces two identical daughter cells
ii. Helps in maintaining uniformity within the species
iii. It is responsible for development of a zygote into adult organism
iv. It is essential for normal growth and development, gives specific shape
v. Acts as a repair mechanism by replacing the old, decayed and dead cells
vi. Production of new individuals in vegetatively propagated crops.
Lec 4 : Cell division – meiosis and its significance

Meiosis : Two successive spindle using divisions which reduce the chromosome number from diploid
to haploid.
Synapsis : Pairing of two homologous chromosomes during zygotene stage of prophase I
Synaptonemal complex: A protein framework which is found between paired chromosomes.
Crossing over: Process of exchange of chromosome segments between non-sister chromatids of
homologous chromosomes.
Chiasma (pl:chiasmata): The point of contact between non-sister chromatids of homologous
chromosomes during crossing over.
Bivalent: The structure formed due to pairing of two homologous chromosomes. The bivalent
consist of four chromatids (tetrad).

MEIOSIS
[Link] in 1905 coined the term meiosis. Cells that undergo meiosis are called meiocytes. This
division occurs in sexually reproducing cells. Meiosis is the process by which haploid gametes or
spores are produced by two successive divisions of diploid nucleus. During meiosis, homologous
chromosomes pair, replicate once and undergo assortment so that each of the four meiotic
products receives one representative of each chromosome. The two nuclear divisions are called
first (meiosis- I) and second meiotic division (meiosis- II).
First meiotic division (meiosis-l):
In meiosis I, the chromosome number is reduced from diploid to haploid. The mechanism consists of
four important phases – prophase l, metaphase I, anaphase I and telophase I.

Prophase I:
The most complex phase of meiosis I is prophase I, it is longer in duration, and consists of five sub
stages – leptotene, zygotene, pachytene, diplotene and diakinesis.
Leptotene is marked by the appearance of the chromosomes as long threads.
Zygotene- homologous chromosomes pair side by side and gene by gene with each other. This
process of lateral association of homologues is called synapsis. When the two homologous
chromosomes consisting of four chromatids are paired (tetrad), this structure is called a bivalent.
In pachytene stage, shortening and thickening of chromosomes takes place. During this stage
crossing over takes place resulting into exchange of portions of homologous [Link]
point of exchange of chromatids during crossing over is called chiasma. Synaptonemal complex
can be seen between synapsed chromosomes.
In diplotene, homologous chromosomes begin to separate, particularly in the region surrounding
the centromere. The sister chromatids remain attached at the centromeric region, at some points
homologous chromosomes remain in close contact, these points are known as chiasmata. Nucleus
decreases in size and nuclear membrane disappears.
Diakinesis, is characterized by shortened chromosomes and the terminalization of chiasmata.
The nucleolus disappear and the nuclear membrane dissolves completely.

Metaphase I:
The homologous chromosomes which are joined through the chiasmata become oriented on the
spindle, with the centromeres of each chromosomes lying towards poles but the ends of
chromosomes towards the equatorial plate.
Anaphase I:
The chromosomes in each bivalent separate at this stage so that homologous pairs disjoin and
migrate towards the opposite poles. As a result, the maternally and paternally derived
homologues are segregated.
Telophase I:
It is a reorganization phase. Nuclear membrane and nucleolus reappear and thus at each pole a
haploid nucleus is formed.

Second Meiotic division (meiosis -II):

It is similar to mitotic division; the sub stages are prophase II, metaphase II, anaphase II and
telophase II. In prophase II, the chromosomes condense, and the centromeres divide. In metaphase
II, a spindle apparatus is organized and the chromosomes become aligned at the equatorial
plate.
In anaphase II, the centromeres migrate to the opposite pole of the spindle, pulling the chromatids
with them. Each of the two cells produced by the first division divides in telophase II, resulting into
formation of four haploid cells. The chromosomes become less condensed and a nuclear
membrane forms.

Significance:
(i) Meiosis is necessary part of the life cycle of sexually reproducing animals and plants as it
helps in restoring the definite number of chromosomes, the characteristic of a species.
(ii) Meiosis is most essential for the completion of life cycle of a plant as it brings a change from
diploid to haploid generation.
(iii) It facilitates segregation and independent assortment of chromosomes and genes.
(iv) The crossing over between homologous chromosomes helps in exchange of genes leading to
formation of new recombinants.
(v) In sexually reproducing species, it is essential for the continuity of generation.
Lec 5: Gametogenesis and syngamy in plants-identical and fraternal twins.

Sporogenesis : Production of microspores and megaspores in anther and ovule.

Spores : Haploid (n) cells produced after meiosis in androecium or gynoecium
Gametogenesis : Production of male and female gametes in microspores and megaspores
Fertilization : Fusion of sperm with egg cell producing a diploid zygote
Zygote: A cell formed by the fertilization of female and male gametes.
Triple fusion : Fusion of another sperm with secondary nucleus leading to the formation of triploid
endosperm.
Double fertilization: Occurrence of syngamy (fertilization) and triple fusion occur in an embryo
sac.
Sporogenesis : It is the production of microspores and megaspores in anther and ovule.
Two types : 1. Micro Sporogenesis 2. Mega Sporogenesis
Gametogenesis : It is the production of male and female gametes in microspores and
megaspores. Two types-1. Micro Gametogenesis 2. Mega Gametogenesis
Funiculus : Stalk of ovule-Ovules are attached to the placenta in the ovary.
Hilum: Point of attachment of body of ovule with funiculus
Raphe : Longitudinal ridge formed by lengthwise fusion of funiculus with body of ovule
Nucellus: Parenchymatous tissue surrounded by integuments.
Integuments: Outer coverings of ovule providing protection to developing embryo
Embryo-sac: Female gametophyte containing egg apparatus
Micropyle: Pore formed by integuments
Chalaza: Place of origin of integuments or basal swollen part of ovary.

Pollination : The transfer of pollen from the male anther to the female stigma

Sporogenesis : 1. Micro Sporogenesis 2. Mega Sporogenesis

Micro Sporogenesis
It occurs in anthers. Anther has the outermost layer (epidermis) below which is
endothecium. Below endothecium, are 1-3 middle layers of parenchyma cells. Cells of
innermost layer radially elongated termed as tapetum which provide nourishment to
developing microspore mother cells (Pollen Mother Cell).
 Each anther has four pollen sacs which contain numerous Pollen Mother Cells (PMC’s) /
Microspore Mother Cell. Nucleus of each PMC undergoes meiosis to produce four haploid
microspores (Tetrad). Four microspores separate & acquire characteristic shape.
Each microspore matures into a pollen grain. Microspore is haploid, uninucleate, with
minute spores. It is generally spherical, can be oval, ellipsoidal, triangular etc in shape.
Cytoplasm covered by outer thick cuticle layer called exine and at some places thin & termed
as germ pores. Inner thin layer is called as intine, composed of pecto-cellulose. Cytoplasm is
rich in starch & unsaturated oils.

Microgametogesis and development of male gametophyte

Microspore is the first cell of gametophytic generation. First mitotic division produces
large vegetative cell or tube cell & small generative cell. Temporary callose wall laid between
two cells dissolves & generative cell comes to lie in tube cell. Two celled pollen grain is ready to
liberate from pollen sacs. Liberated pollen grain falls on stigma. Tube cell comes out of germ
pore and form pollen tube. Generative nucleus undergo second mitosis forms two nuclei &
later on male gametes. Tube nucleus also migrates into pollen tube.
 2. Mega Sporogenesis
The ovule contains the female reproductive cells. Archesporial cell of ovule directly
function as megaspore mother cell (MMC) or divide into outer parietal cell & inner sporogenous
cell. The sporogenous cell behaves as MMC. Formation of megaspore from megaspore mother
cells is megasporogenesis. MMC undergoes meiotic division to form four megaspores.
Three megaspores degenerate & one remain functional.

Megagametogenesis & Development of Female Gametophyte

Functional megaspore develops into female gametophyte (embryosac) through three
mitotic divisions. First, nucleus of functional megaspore divides mitotically to form two nuclei
moving to opposite poles. Two more sequential mitotic divisions produce eight nucleate stage, four
at micropylar end & four at chalazal end.
Egg apparatus consists of egg & two synergids. Synergids have cellular thickenings at
micropylar end called filiform apparatus which helps in absorption & transportation of nutrients.
Three cells at chalazal end called antipodal cells
and large central cell has two polar nuclei.
Pollination: Pollination brings the pollen from anther into the female stigma of a flower. The
pollen is then transferred into the pistil or gynoecium, which contains the stigma, style and ovary.
Its tube cell sticks to the receptive stigma and starts to develop into a pollen tube. The male
germ cell releases two sperm cells that travel to reach the flower’s ovule. While the male cell
grows, the female cell in the ovule develops. One of the sperm cells fuses with the egg to form
the zygote, while the other sperm fuses with polar nuclei to forms an endosperm, which serves
to nourish the growing zygote or seed. After fertilization occurs, the ovary wall becomes a fruit.

Plant embryogenesis is the process that produces a plant embryo from a fertilized ovule by
asymmetric cell division and the differentiation of undifferentiated cells into tissues and organs.
It occurs during seed development, when the single-celled zygote undergoes a programmed
pattern of cell division resulting in a mature embryo. A similar process continues during the plant's
life within the meristems of the stems and roots.
Embryogenesis occurs naturally as a result of sexual fertilization and the formation of the zygotic
embryos. The embryo along with other cells from the mother plant develops into the seed or the
next generation, which, after germination grows into a new plant.
Alternation of generations
Life cycle of most plants has two distinctive generations: a haploid gametophytic (gamete
bearing plant) generation and a diploid sporophytic (spore bearing plant) generation.
Gametophyte produce gametes which unite to form diploid sporophyte which in turn give rise to
spores that develop into haploid gametophytes. This process is referred to as the alternation of
generations.
BIOLOGY OF TWINNING
1. Monozygotic, one egg, or identical twins
2. Dizygotic, two egg, or fraternal twins

Identical twins
Identical twins are derived from a single zygote due to an abnormal division of the embryo
very early in the development. To form identical twins, one fertilised egg (ovum) splits and
develops two babies with exactly the same genetic [Link] twins , therefore, are
always of same sex and have identical genotypes.

Fraternal twins
Fraternal twins are derived from two different zygotes resulting from fertilization of two
different eggs by separate sperms. As a result, such twins have different genotypes, and may be
of different sexes. Twins are more or less equally likely to be female or male.

Difference between Identical and Fraternal Twins

Characters Fraternal Twins Identical Twins

Develop from Two different eggs fertilized by two The splitting of the same fertilized egg
different sperm cells into two

Genetic code Like any other sibling; not identical. Nearly identical

Gender Usually different Always the same

Likelihood About 6 in 1,000 in Japan, up to over Uniform around the world; about 3 in
20 per 1,000 in some parts of Africa. 1,000. Only one-third of all twins in
Two-thirds of all twins in the world the world are identical.
are fraternal.
Characters Fraternal Twins Identical Twins

Blood type May be different Always the same

Causes Hereditary predisposition, certain Not known

fertility drugs, IVF

Appearance As similar as any other sibling Extremely similar, may not be exactly
identical due to environmental factors

Placenta Separate placentas Shared placenta

Fingerprints Different Different

Lec 6: Chromosome structure, chemical composition, nucleosome, centromere,
telomere, euchromatin, heterochromatin, NOR, satellite chromosome, karyotype,
ideogram
Chromosome: Darkly stained nucleoprotein bodies that are visible under microscope in a cell
during cell division.
Haploid number: The gametic chromosome number of a species, represented as n
Diploid number: The somatic chromosome number of a species, represented as 2n.
Basic number: The gametic chromosome number of a true diploid species, represented as x.
Centromere: The region where two sister chromatids of a chromosome “joined together” during
mitotic metaphase (or) the region of chromosome with which the spindle fibres attached during
metaphase. Also called as primary constriction or kinetochore.
Chromatid: One of the two distinct longitudinal sub-units of a chromosome.
Secondary constriction: The narrow or constricted region in a chromosome other than centromere.
Telomere: The terminal region of chromosome on either side.
Chromomere: Linearly arranged bead shaped structures found on the chromosomes.
Chromonema: Thread like coiled structures present in the chromosomes and chromatid during all
stages of mitosis ([Link]).
Matrix: A mass of acromatic material in which chromonemata are embedded.
Karyotype: The characteristic features of chromosomes of a species (or) the number, size and
morphology of the chromosomes of a species.
Idiogram: The diagram which is used to represent karyotype. It is generally depicted in the
descending order of chromosome length.
Euchromatin : Lightly staining regions of chromosomes during interphase. Usually found in the
middle of the chromosome. Genetically active and takes part in transcription.
Heterochromatin : Darkly stained region of chromosome during interphase. Usually inactive in
transcription and found near the centromere and telomere.

Historical background
The Chromosomes were first discovered by Strasburger (1875) and the term “Chromosome” was
coined by Waldeyer in 1888. The word chromosome comes from the Greek word chroma - color
and soma - body due to their property of being very strongly stained by particular dyes.
Chromosomes are darkly stained, rod-shaped, filamentous bodies present in the nucleus, which
become visible under light microscope during cell division. They are the carriers of the gene or
unit of heredity. Their number can be counted easily only during mitotic metaphase.
A chromosome is a structure that occurs within cells and that contains the cell's genetic material.
That genetic material, which determines how an organism develops, is a molecule of DNA. A
molecule of DNA is a very long, coiled structure that contains many identifiable subunits known
as genes. In prokaryotes, the cells without a nucleus, the chromosome is circular DNA. In
eukaryotes, the cells with a distinct nucleus, chromosomes are much more complex in structure.

Chromosome size
In contrast to other cell organelles, the size of chromosomes shows a remarkable variation
depending upon the stages of cell division. During interphase, chromosome is longest & thinnest.
There is a progressive decrease in their length accompanied with an increase in thickness during
prophase. In metaphase, chromosomes are the most easily observed and studied during
metaphase when they are very thick, quite short and well spread in the cell. Anaphase
chromosomes are smallest. Therefore, chromosomes measurements are generally taken during
mitotic metaphase.
The size of the chromosomes in mitotic phase of animal and plants sp generally varies
between 0.5 µ and 32 µ in length, and between 0.2 µ and 3.0 µ in diameter. The longest
metaphase chromosomes found in Trillium is 32 µ in length. In general, plants have longer
chromosomes than animal and species having lower chromosome numbers have long chromosomes
than those having higher chromosome numbers. Among plants, dicots in general, have a higher
number of chromosomes than monocots. Chromosomes are longer in monocot than dicots.

Structure of a chromosome
The chromosomes are usually studied in the cells of root tip during mitotic metaphase under light
microscope. It consists of the following parts.

1. Centromere
Each chromosome has a constriction point called the centromere, which divides the chromosome
into two sections, or “arms.” The short arm of the chromosome is labelled the “p arm.” The long
arm of the chromosome is labelled the “q arm.” It is also termed as Primary constriction or
kinetochore. When chromosomes are stained they typically show a dark-stained region. It is the
region to which microtubules attach during cell division. The important functions are
i. Orientation of metaphase chromosomes at the equatorial plate
ii. Movement of chromosomes during anaphase
iii. During mitotic anaphase and meiotic anaphase II, the centromere divides so that the
chromatids can migrate to opposite poles of the cell.
iv. Gives the chromosome its characteristic shape (terminal, sub-terminal or median).
2. Chromatid
One of the two distinct longitudinal subunits of a chromosome is called as chromatid. Two
chromatids of chromosomes are held together by centromere. These get separated during
anaphase. The chromatids arise due to replication during S phase. The chromatids that arise from
the same chromosome that is still attached to a common centromere is known as sister chromatids
and the chromatids of homologous chromosomes that involve in crossing over during anaphase I of
meiosis is called as non-sister chromatids. After centromere division, each chromatid becomes a
chromosome.

3. Secondary constriction
It is the constricted or narrow region other than centromere present either in short or long arm.
Position of these constrictions are constant, hence these constrictions are useful in identifying
particular chromosomes in a set. Number varies from species to species. It is distinguished from
centromere, because chromosome bends only at the position of centromere during anaphase.

4. Teleomere
The terminal region of a chromosome on either end of a chromosome is known as telomeres.
It required for the replication and stability of the chromosome. When telomeres are damaged or
removed due to chromosome breakage, the damaged chromosome ends can readily fuse or unite
with broken ends of other chromosome. Thus it is generally accepted that structural integrity and
individuality of chromosomes is maintained due to telomeres. The telomere of one chromosome
cannot unite with the telomere of other chromosome due to polarity effect.

5. Satellite
Sometimes the chromosomes bear round elongated or knob like appendages known as satellites.
The satellites remain connected with the rest of the chromosome by a thin chromatin filament. The
chromosomes with the satellite are sat chromosomes. The shape and size of the satellite remain
constant.

6. Nucleolar Organizing Region (NOR)

The chromosome having secondary constriction is known as nucleolus organizer region because it
produces nucleolus associated with secondary constriction. Nucleolus is always associated with
secondary constriction. In human beings NOR are located in the secondary constrictions of
chromosomes 13,14,15,21 and 22, all of which are acrocentric and have satellites.

A. Nucleolus organizer B. Chromosome C. Nucleolus

7. Chromomeres
Serially arranged small bead shaped, heavily staining bodies resulting from local coiling of a
continuous DNA thread. Distribution of chromomeres is constant. It is especially clear in the
polytene chromosomes. It represents unit of DNA replication, chromosome coiling, RNA synthesis
and RNA processing.

8. Chromonema
Thread like coiled structures present in a chromatid during all stages of mitosis. During mitotic
prophase, chromosomal material visible as very thin filaments called chromonemata. It represents
a chromatid in the early stages of condensation. Chromatid may contain one or
more chromonema. It is associated with size of chromosomes, duplication of chromosomes and
forms the gene bearing portion of chromosomes.

9. Matrix
Each chromosome is bounded by a membrane called pellicle. It is very thin and is formed of
achromatic substance. This membrane encloses a jelly-like substance which is called matrix. It is a
mass of acromatic material in which chromonemata are embedded. Matrix and pellicle are non-
genetic material.

Composition of chromosome
The material in which the chromosomes are composed is called chromatin. Term chromatin
introduced by [Link] in 1879. It is classified into two classes, viz., heterochromatin (darkly
packed) and euchromatin (lightly packed chromatin). Euchromatin comprises the most active
portion of the genome within the cell nucleus. Generally heterochromatin found in centromeric &
telomeric regions.
Heterochromatin is further divided into 2 groups:
a) Constitutive heterochromatin: Present in all cells at identical positions on both homologous
chromosomes of a pair.
b) Facultative heterochromatin: It varies in state in different cell types, at different stage or
sometimes, from homologous chromosome to another. e.g., Barr body, an inactivated X
chromosome in somatic cells of mammalian female (XX).
Euchromatin : It is the active region of the chromosome, involved in transcription. It contains
structural genes which replicate and transcribe during G1 and S phase of interphase.
 Heterochromatin Euchromatin

Represent darkly stained regions Lightly stained regions

High content of repetitive DNA. Contains lot of active genes
Contains few inactive genes
Covers small region of chromosome Larger region of chromosome
Usually found near centromere & Found in the middle of chromosome between
telomere centromere & telomere
Late replicating Normal replicating
Usually no active part in transcription Plays active roles in transcription and 8.3-

Organization of chromosomes
1. The folded fiber model (DuPraw,1965)
2. Nucleosome-Solenoid model (Kornberg & Thomas,1974) –widely accepted

Organization of DNA into a chromosome

Eukaryotic chromosome contain single giant DNA molecule of DNA. DNA wrapped
around an octamer of small basic proteins called histones. The histone proteins have basic
properties and have significant role in controlling or regulating the functions of chromosomal DNA.
Histone proteins are rich in arginine & lysine. It is of 5 types (H1, H2A, H2B, H3, and H4).This
DNA-histone complex is called nucelosome. Nucleosome is the simplest packaging structure of
DNA that is found in all eukaryotic chromosomes. Each nucleosome contains a highly conserved
core consisting of 146 nucleotide pair long segment (core DNA) wrapped around a histone
octamer. Each octamer contains two copies of histone proteins (H2A, H2B, H3, and H4) these are
known as core histones.

The super coiled nucleosome fibre is known as solenoid. Each solenoid is linked to another
by linker or spacer DNA of four bases with one molecule of H1 protein. According to
Nucleosome-Solenoid model, a very long molecule of DNA (146bp) is packed into a single
nucleosome and several units of nucleosome constitute solenoid fibre (chromatin fibre). This
chromatin fibre is 300 Å in diameter which is visible under electron microscope. At metaphase,
these fibres are in turn coiled into highly condensed structures that can be seen under light
microscope. The third level of condensation involves a chromosomal scaffold that is composed
of non-histone proteins.
Chemical composition of chromosome
Eukaryotic chromosomes composed of DNA, RNA, histone and non-histone proteins and certain
metallic ions. Important enzymatic proteins are phosphoproteins, DNA polymerase, RNA-
polymerase, DPN-pryrophosphorylase and nucleoside triphosphate. The metal ions Ca+ and
Mg+ are supposed to maintain the oragnization of chromosomes intact.

Karyotype
Karyotype is the phenotypic appearance of chromosomes. It represents the number, size
and morphology of chromosome set of a cell or species. It is the group of characteristics that
identifies a particular set of chromosomes. It specifies number, position, size, degree & distribution
of heterochromatin. Represented by arranging the somatic chromosome complements according to
their length. Generally, karyotype is represented by arranging the chromosomes in descending
order of size, keeping their centromeres in the same line. It is identical for particular species.
Idiogram or idiotype
Diagramatic representation of a karyotype (or morphological features of the chromosome) of a
species is called idiogram (Gr.,idios = distinctive; gramma = something written). In an ideogram,
the chromosomes of a haploid set of an organism are ordered in a series of decreasing size.
Arrangement of chromosome can be improved by determining centromeric index, which is the
ratio of the lengths of the long and short arms of the chromosome.

Uses of karyotype
1. It helps in the understanding of evolutionary process.
2. It can be used to study chromosomal aberrations, cellular function, taxonomic relationships.
3. It also suggests primitive or advanced features of an organism
4. A karyotype exhibiting large differences in smallest and largest chromosomes of the set and
containing fewer metacentric chromosomes is called asymmetric karyotype. When compared
to symmetric, asymmetric karyotype is considered to be relatively advanced feature.

Banding techniques
Banding techniques have been developed to study the karyotype in plants and animals.

C Banding: C banding demonstrates constitutive heterochromatin presence around the

centromere, telomere and in the nucleolar organizer region.
G banding: It is useful to distinguish between euchromatic (stains lightly) and heterochromatic
regions (stain darkly) of a chromosome.
R (Reverse) banding: R banding is just the reverse of G banding in the lightly stained G bands
become darkly stained.
Q banding : The dye Quinacrine binds preferentially to DNA regions rich in A-T base pair.
Lecture 7: Types of chromosomes based on position of centromere, based
on structure and function: normal and special chromosomes - polytene,
lampbrush, based on the role in sex determination: autosomes and
allosomes, Other types of chromosomes - B, ring and isochromosomes.

Metacentric chromosome: A chromosome in which centromere is located in the

middle portion. Such chromosome assumes V shape at anaphase.
Sub-metacentric chromosome: A chromosome in which centromere has sub-median
position, which assumes J shape or L shape at anaphase.
Acrocentric chromosome: A chromosome in which centromere is located very near
to one end or has sub-terminal position which assumes J shape at anaphase.
Telocentric chromosome: A chromosome in which centromere is located at one end.
Holokinetic chromosome: A chromosome with diffused centromere.
Monocentric chromosome: A chromosome with one centromere.
Dicentric chromosome: A chromosome with two centromeres.
Rod shaped chromosome: A chromosome which assumes the shape of rod at
anaphase.
Linear chromosome: A chromosome having both ends free. Found in eukaryotes.
Circular chromosome: A chromosome with circular shape. Found in bacteria and
viruses.
Lampbrush chromosome: A chromosome having lampbrush appearance.
Polytene chromosome: The multiple replicates of the same chromosome holding
together in a parallel fashion resulting in very thick structure. Also called as giant or
salivary gland chromosome.
Bands: Darkly staining strips found on polytene chromosome
Puffs: The swollen regions found on polytene chromosome. Also called as ―Balbiani
rings‖
Isochromosome: A chromosome with two identical arms.
Ring chromosome: A ring chromosome is a chromosome whose arms have fused
together to form a ring. Usually found in prokaryotes such as bacteria and viruses.
Chromosome banding: The differentially stained regions of chromosomes as a result
of treatment with various dyes, visible under light or fluorescence microscope.

Type of Chromosomes
1. Based on the number of centromeres

The number and position of centromeres is variable, but is definite in a specific

chromosome of all the cells and in all the individuals of the same species. Thus,
according to the number of the centromere the eukaryotic chromosomes may be
acentric (without any centromere), mono centric (with one centromere), dicentric
(with two centromeres) or polycentric (with more than two centromeres). In a
number of animals, especially in insects of the order Hemiptera and few
monocotyledonous plants, the kinetic activity is distributed over the entire
chromosome and such chromosomes are called Holokinetic chromosomes (Sybenga,
1972).

2. Based on the position of the centromere

According to the position of the centromere, the eukaryotic chromosomes may be
rod-shaped (telocentric and acrocentric), J-shaped (submetacentric) and V-shaped
(metacentric). During the cell divisions the microtubules of the spindle are get
attached with the chromosomal centromeres and move them towards the opposite
poles of cell.

A. Centromere
Acrocentric Telocentric Metacentric Sub metacentric
3. Based on Structure and appearance
Linear : Linear structure or having both ends free; found in eukaryotes
Circular : Found in bacteria and virus
4. Based on function
Allosomes (sex chromosomes): Differ in morphology and number in male and
female sex and contain sex determining genes. E.g.: X and Y chromosomes in
human beings and Drosophila.
Autosomes : Do not differ in morphology and number in male and female sex. and
contain genes which determine somatic characters of individuals and represented
by letter ‗A‘.
5. Based on essentiality
A chromosome: Normal members and essential for normal growth and
development
B chromosome: Found in addition to normal chromosome. It is also called as
accessory, supernumerary or extra chromosomes. Not essential for normal
growth and development.

6. Based on Structure and Function

Normal chromosome : Normal structure (shape and size) and function
Special chromosome : Significantly differ in structure and function;eg,
Lampbrush, polytene and B-chromosomes
Special Types of Chromosomes
The eukaryotes besides possessing the usual type of chromosomes in their body cells,
contain some unusual and special types of chromosomes in some body cells or at
some particular stage of their life cycle. The special types of eukaryotic
chromosomes are following:
1. Polytene chromosomes
2. Lampbrush chromosomes
3. Isochromosome
4. Ring chromosomes
5. B-chromosomes / accessory chromosomes

1. Polytene chromosomes
It was discovered by E. G. Balbiani (1881) in Dipteran salivary gland. The nuclei of the
salivary gland cells of the larvae of Dipterans like Drosophila have unusually long
and wide chromosomes, 100 or 200 times in size of the chromosomes. Found in other
tissues viz., gut epithelium, Malphigian tubules of some Diptera. It is also known as
polytene chromosomes or multistranded chromosomes or giant chromosomes. The
total length of Drosophila melanogaster polytene chromosome is about 2000µ.

The polytene Chromosomes of the salivary gland cells of D. melanogaster contain

1000 to 2000 chromosomes. The numerous strands of these chromosomes are
produced due to repeated replication of the paired chromosomes without any nuclear
or cell division (endomitosis). The number of strands (chromatids) in a chromosome
doubles after every round of DNA replication. Giant chromosomes of Drosophila have
about 1,024 strands. In Chironomous, a giant chromosome has about 4,096 strands.
Giant chromosome has several dark staining regions called ―bands‖. Bands separated
by relatively light or non-staining ―interband‖ regions. The bands in Drosophila giant
chromosome are visible without staining. The bands of giant chromosomes are formed
as a result of stacking over one another the chromomeres of all strands present in
them. Some of these bands are as thick as 0.5µ. About 25,000 base-pairs are
estimated for each band. Since chromatin fibers are highly coiled in band region of
chromosomes, they stain deeply. The dark bands are comparable with the
chromomeres and are disc-shaped structures occupying the whole diameter of
chromosome. They contain [Link] the other hand, the chromatin fibers in
the interband regions are fully extended, as a result these regions take up very light
stain. The inter bands are fibrillar and composed of heterochromatin. In Drosophila,
the location of many genes is correlated with specific bands in the connected
chromosomes. Interband region do not have functional genes.

The polytene chromosomes of dipteran larval salivary glands had specific areas (sets
of bands) which is enlarged ("puff"/ Balbiani rings). Puffs are produced due to
uncoiling of chromatin fibers present in the concerned chromomeres. Such puffs
change locations as development [Link] temporal puffing indicates changes in
gene activity and involves several processes such as the accumulation of acidic
proteins, despiralization of DNA, formation of chromonemal loops, synthesis of mRNA
(messenger RNA) and storage (accumulation) of newly synthesized mRNA around the
Balbiani rings.

[Link] chromosomes
It was given this name because it is similar in appearance to the brushes used to clean
lamp chimneys. It was first observed by Flemming in 1882. The name lampbrush was
given by Ruckert in [Link] diplotene stage of meiosis, the yolk rich oocytes of
vertebrates contain the nuclei with many lamp brush shaped chromosomes of
exceptionally large sizes. The lampbrush chromosomes are formed during the active
synthesis of mRNA molecules for the future use by the egg during cleavage when no
synthesis of mRNA molecules is possible due to active involvement of chromosomes in
the mitotic cell division.
Each lampbrush chromosome contains
a central axial region, where the two
chromatids are highly condensed. It
has several chromomeres distributed
over its length. From each
chromomere, a pair of loops emerges
in the opposite directions vertical to
the main chromosomal axis. One loop
represents one chromatid, i.e., one DNA
molecule. The pairs of loops are
produced due to uncoiling of the two
chromatin fibers present in a highly
coiled state in the chromomeres. The
size of the loop ranging from 9.5 µm to
200 µm.
Lampbrush chromosomes are up to 800 µm long; thus they provide very favorable
material for cytological studies. One end of each loop is thinner (thin end) than the
other end (thick end). There is extensive RNA synthesis at the thin end of the loops,
while there is little or no RNA synthesis at the thick end.

3. Isochromosome

Isochromosome are having identical arms and

identical genes. The arms are mirror images of
each other. Transverse division of centromere,
leads to the formation of isochromosome.
Extensively studied in maize.
At meiosis isochromosomes pair in three
different ways.
i. Internal pairing : the two arms of the
isochromosmes pair with each other.
ii. Fraternal pairing : one or both of the
arms of the isochromosomes pair with a
homologous arm of another
chromosome.
iii. Normal pairing: the isochromosome
pairs with another one just like it.
4. Ring chromosomes
i. The chromosomes of higher organisms with normal chromosomes do not form
rings because they are believed to have telomeres on each end. Telomeres
prevent the union of chromosome arms into ring formation. When the ends of
a chromosomes are lost, enabling the arms to fuse together.
ii. A chromosome can form a ring chromosome by fusion of the raw ends only if it
has two terminal deletions producing centric segment with two raw ends and
two acentric fragements.
iii. Discovered by Lilian Vaughan Morgan in 1926.

iv. A ring chromosome is denoted by the symbol „r‟ in human genetics or „R‟ in
Drosophila genetics. In Prokaryotes, ring shaped chromosomes referred as
genophores. Ring shape chromosomes are found Humans, Drosophila and
plants.
v. A ring chromosome lacks the genetic information that was carried by the
terminally deleted fragments.
vi. Ring chromosomes form by mutagens, but they may also arise spontaneously
during development
vii. They are meiotically unstable and they are associated with several syndromes.
viii. Human genetic disorders can because by spontaneous ring chromosome
formation. Ring formation of an X-chromosomes causes turner syndrome.
ix. Symptoms seen in patients carrying ring chromosomes is due to the deletion
of genes in the telomeric regions, rather than by the formation of a ring
structure itself.

5. B-chromosomes
Many plant (maize, rye) and animal (such as insects and small mammals) species,
besides having autosomes (A-chromosomes) and sex-chromosomes possess a special
category of chromosomes called B-chromosomes without obvious genetic function.
These B-chromosomes (also called supernumerary chromosomes or accessory
chromosomes or extra chromosome) usually have a normal structure and can be
predominantly, heterochromatic (many insects, maize, etc.) or pro-dominantly
euchromatic (rye). They are not essential for normal growth and development. It was
discovered in maize by Randolph and Longley (1827). It may found in organisms as
extra chromosomes over and above the diploid or polyploidy chromosome
complements.
In animals, the B-chromosomes disappear from the non-reproductive (somatic) tissue
and are maintained only in the cell-lines that lead to the reproductive organs.
B-chromosomes have negative consequences for the organism, as they have
deleterious effect because of abnormal crossing over during meiosis. In animals, B-
chromosomes occur more frequently in females and the basis is non-disjunction. The
non-disjunction of B-chromosomes of rye plant is found to be caused due to the
presence of a heterochromatic knob at the end of long arm of B-chromosome.
The ‗B‘ chromosomes are smaller than ‗A‘ chromosome and genetically inert. It is not
homologous with ‗A‘ chromosome. It suppresses the vigour and fertility. In maize,
they adversely affect, development and fertility only when occur, in large amount.

Genetic Significance of Chromosomes

The chromosomes are considered as the organs of heredity because of the following
reasons:
(i) They form the only link between two generations.
(ii) A diploid chromosome set consists of two morphologically similar (except the X
and Y sex chromosomes) sets, one is derived from the mother and another from the
father at fertilization.
(iii) The genetic material, DNA or RNA is localized in the chromosome and its contents
are relatively constant from one generation to the next.
(iv) The chromosomes maintain and replicate the genetic information contained in
their DNA molecule. This information is transcribed at the right time in proper
sequence into the specific types of RNA molecules which directs the synthesis of
different types of proteins to form a body form like the parents.
Lec 8. Chromosomal aberration: Variation in chromosome structure – deletion,
duplication, inversion and translocation – genetic and cytological implications.
Definitions
Structural chromosomal change: Any change which alters the normal structure of a
chromosome.
Restitution: Union of broken segments which restores original gene sequence.
Deletion: Loss of segment from a chromosome, also called as deficiency. It is two
types viz., Terminal and Interstitial.
Terminal deletion: Loss of either terminal segment of a chromosome.
Interstitial deletion: Loss of intercalary segment of a chromosome.
Duplication: Occurrence of a segment twice in the same chromosome. It is of four
types viz.,Tandem, reverse tandem, displaced and reverse displaced.
Tandem: Duplication with normal sequence (similar to original sequence) of genes.
Reverse tandem: Duplication with reverse sequence of genes as compared to the
original segment.
Displaced: Presence of duplication away from the original segment but on the same
arm of the chromosome.
Reverse displaced: Presence of duplication away from the original segment but on
the other arm of the chromosome.
Inversion: Segment of chromosome is oriented in the reverse order. It is of two types
viz., paracentric and pericentric
Paracentric inversion: Inversion in which centromere is not involved.
Paricentric inversion: Inversion in which centromere is involved.
Translocation: One way or reciprocal exchange of segments between non-homologous
chromosome. It is of three types-simple, shift and reciprocal.
Simple Translocation: Transfer of terminal segment from one chromosome to the end
of a non-homologous chromosome.
Shifts: Transfer of intercalary segment from one chromosome to the intercalary
position in a non-homologous chromosome.
Alternate segregation: At anaphase, movement of two normal chromosomes (N1 and
N2) towards one pole and that of translocated chromosomes (T1 and T2) to another
pole.
Adjacent 1 segregation: Segregation of one normal chromosome with one
translocated chromosome (N1+ T2 and N2+T1).
Adjacent 2 segregation: Segregation of T1N1 to one pole and T2N2 to another pole.
Chromosomal aberration

The chromosomes of a genome are distinct from each other in their morphology
and or gene content. Each chromosome of a genome contains definite number of
genes arranged in definite sequence. Homologous chromosomes contain identical
number of genes in same sequence. Chromosomal aberration involves variation in
chromosomal number or structure occurs.

Chromosomal aberration grouped into two broad classes:

1. Structural aberration
2. Numerical aberration

Structural Chromosome Aberration

In this category, aberration alters the chromosome structure but do not involve a
change in chromosome number. These aberrations involve rearrangement through
loss, gain or reallocation of chromosomal segments.

• Chromosomal aberrations which alter the chromosomal structure i.e. the no. of
genes, the sequence present in the chromosome(s) and do not involve a change
in chromosome number.
• Chromosome structure variations result from chromosome breakage.
• Broken chromosomes tend to re-join;
• Chromosome breakage is caused by X-rays, various chemicals, and can also
occur spontaneously.

Depending upon number of breaks, their location & pattern of joining of broken ends,
chromosomal aberrations are of 4 major types.

Intra-chromosomal aberrations:

1. Deletion or Deficiency
2. Duplication or Repeat
3. Inversion
Inter-chromosomal aberrations:
[Link]
A) Deletion or Deficiency:
Deletion or deficiency as the name
suggests there is a loss of segment of
chromosome. After break the part
without centromere is lost. On the other
hand, the part attached to the
centromere acts as deficient
chromosome. Bridges (1917) for the first
time observed deficiency in the X
chromosome of Drosophila.

Two types of deletions are found:

Terminal deletion:
A single break occurs near the end of the
chromosome.
Interstitial deletion:
Two chromosome breaks occur and reunites but
the part is lost. Deletions are detected at the
time of homologous pairing. If a part of chromosome is missing then the other
chromosome also has to omit it in the form of bulging in order to make synapse. e.g.,
if a chromosome has 1, 2, 3, 4 genes. The part 2 is missing from one chromosome
leaving, 1, 3, 4. The other homologous chromosome at the time of synapse bulge out
or form loop at position [Link] the missing segment is of physiological importance the
individual will not survive.

Based on the involvement of either single or both members of homologous

chromosomes, deletions are

1. Deletion homozygote

Both members of homologous chromosomes have deletions at corresponding

regions. Hence, normal chromosome pairing occurs at pachytene. Homozygous
deletions are generally lethal.

2. Deletion heteroygote

One member of homologous chromosomes having deletions and the other

member of homologous chromosomes does not have deletions. Normal
chromosome forms a loop if the deletion is intercalary and does not form
deletion loop in terminal deletion.
Genetics and cytological effects of deletion

i. Organisms with homozygous deletion do not survive to an adult stage because a

complete set of genes is lacking (lethal effect).
ii. Small deficiencies, if present in heterozygous condition (deficiency
heterozygote) can be tolerated by the organisms. In such individuals during
pachytene stage of meiosis the unpaired segment of the normal chromosome of
an intercalary deletion heterozygote produces a characteristic loop in a
bivalent.
iii. Individual heterozygote for a deficiency contains only one copy of deleted
segment (hemizygous state). Eg. When an organism heterozygous for a pair
of alleles Aa loses a portion bearing the dominant allele ‗A‘, the recessive
allele in the hemizygous condition will get expressed phenotypically. This
phenomenon is known as pseudo-dominance. This principle of pseudo-
dominance utilized for location of genes on specific chromosome in Drosophila
and maize. Thus deletions are important cytological tools for mapping genes.

iv. Deletion plays an important role in species formation and creating variability
through chromosomal mutations.
v. Crossing over is completely absent in the deleted region of a chromosome in
deletion heterozygote.
vi. Chromosome with deletion can never revert back to normal condition. Hence,
karyotype of the individual is changed.
vii. In plants, deletion causes pollen and embryo sac abortion. Female gamete
able to withstand deletion to some extent than pollen grain.
viii. In human, deletion of chromosome 5 results in cri-du-chat syndrome (cat cry
syndrome), children cry like cat, they have small head and are mentally
retarded. Other characteristics are microcephaly (small head), broad face and
saddle nose. Cri-du-chat patients die in infancy or early childhood.
viii. Another human disorder that is associated with a chromosome deletion is
chronic myelocytic leukemia. A deletion of chromosome 21 was called
―Philadelphia‖ (Ph‘) chromosome.
B. Duplication
The presence of an additional chromosome segment, as compared to that normally
present. Duplication was first discovered in Drosophila by C.B. Bridges in 1919. Four
types of duplication:
1. Tandem duplication
2. Reverse tandem duplication
3. Displaced duplication
4. Translocation duplication
Tandem duplication
The extra chromosome segment may be
located immediately after the normal
segment in precisely the same orientation
Reverse tandem
When the gene sequence in the extra segment
of a tandem in the reverse order i.e, inverted
Displaced tandem
The extra segment may be located in the
same chromosome but away from the normal
segment from its original location but on the
same arm (homobrachial displacement) or on
the other arm (heterobrachial displacement).
Transposition
When the segment is duplicated on the non
homologous chromosome it is called
transposition.

Duplication-Origin

Duplication arises due to unequal crossing over between non-sister chromatids. This
give rise to two types of chromatids viz., one of the two homologous ends up with a
deficiency, while the other has a duplication for the concerned segment. e.g.,
duplication of the band 16A of X chromosome of Drosophila (Bar eye) due to unequal
crossing over between the two normal X chromosomes of female.
Genetic and Cytological Effect

 Duplications are not lethal to the individual. They do not reduce viability of
an individual.
 Results in gene redundancy.
 Crossing over is suppressed in the duplicated region due to lack of
corresponding duplicated segment
 During meiotic pairing of heterozygotes, the chromosome with duplicated
segment forms a loop.
 Duplication leads to reduction in pollen fertility
 Duplication leads to addition of genes in a population, plays role in
evolution.
 Position effect: Altered phenotype due to duplication and relocation of
chromosomal segment. [Link]-Eye Phenotype in Drosophila. This is due to
unequal crossing over between the two normal X chromosomes of female.

1. Normal oval shape-Single dose of 16A segment of X chromosome

2. A fly homozygous for Bar eye phenotype with four 16A segments, two in
each X chromosome produces reduced number of facets.
3. A fly heterozygous for ultra Bar (Double bar) also has also four 16A
segments- one in normal and three in duplicated Chromosome. Flies
homozygous for ultra Bar produced smaller size eyes. Ultra-bar (or double-
bar) is a trait in which flies have even fewer facets than the bar homozygote.
This is called dosage effect.
(C) Inversions:

When a segment of chromosome is oriented in the reverse direction, such segment

said to be inverted and the phenomenon is termed as inversion.
The existence of inversion was first detected by Strutevant and Plunkett in 1926.
Inversion occur when parts of chromosomes become detached , turn through 1800 and
are reinserted in such a way that the genes are in reversed order.

Inversion arises by the formation of loops on a chromosome.

Breaks may occur at the point of intersection of the loops (Fig.
43.13). Reunion of the broken ends takes place in a new
combination, and inverts. Inversion heterozygotes are formed
by loops and bulges in pairs.

Types of Inversion:
Paracentric inversion: The inverted segment does not include
centromere.
Pericentric inversion: Inverted segments include centromere

Paracentric or homobranchial inversions:

Inversion homozygote: When both members of

homologous pair have similar type of inversion.
Meiosis is normal in inversion homozygote

Inversion heterozygote: One chromosome of

homologous pair has inversion. Inversion loop is
formed during synapsis. Chromosome carrying the
inverted segment form an inverted loop (twisted
loop), the normal chromosome form a semi-
circular loop over the inverted loop.
A single crossing over inverted region will result into
formation of a dicentric chromosome (with 2
centromeres) and an acentric chromosome (with no
centromere). Of the remaining 2 chromatids one will
be normal and the other will carry inversion. The
dicentric chromatid and acentric chromatid will be observed at anaphase I in the form
of a bridge and a fragment. Double crossover shows deficiencies and duplication
(Fig. 43.15) giving rise to variations in anaphase I configurations. In maize, dicentric
bridge break at random point and chromatids produced are included into gametes.
Since the chromatids produced have duplication and deficiencies, male gametes show
pollen sterility. The egg cells function normally.

Pericetric or heterobranchial inversion

In pericentric inversion, centromere is in the inverted segments and break occurs in
both arms of a chromosome. In pericentric inversion, if two breaks are not situated
equidistant from the centromere, a change in shape of chromosome results. A
metacentric chromosome may become submetacentric and vice versa.

In pericentric heterozygote, synapsis occur by forming inverted loop by inverted

chromosome and a semi-circular loop by normal chromosome. Single cross over
within the inversion loop produces four monocentric chromatids. Two cross over
chromatids have deficiencies and duplications.; one normal parental and one
inverted parental [Link] paracentric inversion, no dicentric bridge or
acentric fragment will be observed at anaphase I. Male gametes carrying crossover
products are inviable and gametes carrying non-crossover chromosomes are viable.

In natural populations, pericentric inversions are much less frequent than

paracentric inversions. Paracentric inversions very frequent in natural populations
of Drosophila.

Genetic and Cytological effect

1. Inversion lead to the production of duplications and deficiencies

2. Paracentric inversion produces dicentric and acentric chromosomes
 3. In inversion heterozygote, chromosome carrying the inverted segment form a
inverted loop, the normal chromosome form a semi-circular loop over the
inverted loop.
4. Cross over in inversion loop leads to production of cross over chromatids
with duplication and deficiencies results in 50% sterility
5. Inversion acts as cross over suppressor
6. Heterozygosity will be maintained generation to generation.
7. Crossing over in inverted loop leads to change in karyotype by shifting the
position of centromere. This leads to the evolution of new species.

(D) Translocation
Transfer of a section of one chromosome to non-homologous chromosome is known as
translocation. It also includes exchange of segments between non homologous parts of
a pair of chromosomes. The segment is neither lost nor added it is just exchanged.

Three types:
1. Simple translocation (unbalanced)
2. Shift translocation
3. Reciprocal translocation (balanced)
Types of translocation:
(a) Simple translocation: Terminal segment of a
chromosome is integrated at one end of a non-
homologous region.
(b) Shift or intercalary translocation:
In shift, an intercalary segment of a chromosome is
integrated in a non-homologous [Link]
type of translocation involving 3 breaks so that a two
break section of one chromosome is inserted within the
break produced in a non homologous chromosome.
(c) Reciprocal translocation:
This involves mutual exchange of chromosome
segments between two non-homologous chromosomes
exchange. Frequently observed translocation where
single break in two homologous chromosomes.
Translocation homozygote:
Both the chromosomes of two pair involved in
[Link] homologues of each of two
translocated chromosomes are [Link] is
normal as that of normal chromosome.
Translocation heterozygote:
Only one chromosome from each pair of two
homologues involved in reciprocal [Link]
remaining chromosome of each pair is normal
chromosome. Most frequent type of translocation.
This type of translocation will not have normal
pairing, but a cross-shaped configuration involving
all four chromosomes observed at pachytene. This
cross shaped configuration include all four associate
chromosomes, with each member of the group
partially homologous to other chromosome in the
[Link]-shaped configuration is effected to have
pairing of all homologous segments of two pairs of
[Link] cross opens out into a ring as
chiasma terminalizes or chain of four chromosomes at
anaphase and chromosomes segregates in 3 different
ways.

Meiotic segregation can occur in one of three ways

1. Alternate segregation
 Two normal chromosomes (N1 & N2) moves towards one pole & two
translocated chromosome (T1 & T2) to opposite pole.
 Adjacent chromosome moves to opposite poles. This is possible by formation
of figure eight.
 All gametes receive full complement of genes & give rise to viable
individual.
 Half of the gametes have normal chromosome and half of the gametes have
balanced translocation chromosome. Leads to balanced gametes.
2. Adjacent-1 segregation
 Segregation of one normal chromosome with one translocated chromosome
which is a non-homologue (T1+N2 and T2+N1).
 This segregation produces non-functional sterile gametes due to the presence
of translocated chromosome with duplications and deletions.
 Leads to unbalanced gametes
3. Adjacent-2 segregation
 Adjacent homologous chromosomes ( N1 + T1) move to one pole and other
homologous chromosomes (T2+N2) move to another pole.
 This is unbalanced translocation with duplications and deletions
 Leads to non-functional or sterile gametes

Genetic and cytological effect

• In the translocation heterozygote, pairing of homologous chromosomal

segments is effected by cross-shaped configuration.
• It leads to duplication and deletion of genes, resulting in the production of
inviable gametes, causing pollen and ovule sterility.
• It alter the chromosome size, number and karyotype, thus play a role in
formation of species.
 Translocation useful in locating the position of genes, centromeres.
• Translocation alter the linkage relationships of genes
• It suppresses crossing over due to competition in pairing.
• Phenotypic expression of genes is modified when it is translocated in to a new
position in the genome. eg. Down‘s syndrome (Mongolism) due to Robertsonian
Translocations which involves the fusion of long arms of acrocentric
chromosomes. 95% of Down Syndrome individuals are a result of Trisomy 21.
However, there is second cause of Down Syndrome caused by a Robertsonian
translocations that is heritable.
Lec 9: Chromosomal aberration: Variation in chromosome number-
euploid,aneuploid, types of aneuploids, Klinefelter syndrome & Turner syndrome

Genome: A basic set of chromosomes in which each type of chromosome is

represented only once.
Basic number: The gametic chromosome number of a true diploid species.
Euploidy: A change in chromosome number which involves entire set of chromosomes
(genome) .
Haploid: An individual with half of somatic chromosome number.
Monoploidy: An Individual with basic chromosome number of a species.
Euhaploid: Haploid which develops from a euploid species.
Monohaploid: Haploid which develops from a normal diploid species.
Polyhaploid: Haploid which develops from a polyploid species.
Allohaploid:Haploid which develops from allopolyploid species.
Autohaploid: Haploid which develops from autopolyploid species.
Aneuploidy: The change in chromosome number which involves one or few
chromosome of the genome.
Hypoploid: An individual lacking one or two chromosomes from the diploid
complements. Such condition is known as hypoploidy.
Monosomic: An individual lacking one chromosome from a diplod set (2n-1).
Nullisomic: An individual lacking one pair of chromosome from a diplod set (2n-2).
Hyperploidy: Addition of one or two chromosomes to one pair or to two different
pairs. It includes trisomics and tetrasomics.
Polysomics: An individual having one or two chromosomes in the diplod set.
Trisomics: Addition of one chromosome to one pair in the diplod set.
Primary Trisomics: The trisomics in which the additional chromosome is a normal
one.
Secondary Trisomics: The trisomics in which the additional chromosome is an
isochromosome.
Tertiary Trisomic: A trisomic having additional chromosome as translocated one.
Tetrasomic : An individual having addition of two chromosomes to one or two
different pairs.
Simple tetrasomic : An individual having addition of two chromosomes to one pairs
(2n+2).
Double tetrasomic : An individual having addition of two chromosomes to two
different pairs (2n+2+2).
 Most plants and animals are diploid and their somatic cells, the chromosomes
are found in pairs. Chromosome number in a species remain constant over generations
which results in constancy of characters. The gametes produced as a result of meiosis
possess half of the chromosomes of somatic cells. Fusion of gametes via fertilization
brings together homologous chromosomes from the two parents and restores the
diploid number.

The somatic cells of a diploid organism contain two sets of homologous

chromosomes (2n number of chromosomes) or in other words, two copies of the same
genome are present. But their gametes contain a single genome i.e. haploid number
of chromosomes (n). An individual carrying the gametic chromosome number, n, is
known as haploid. A monoploid, on the other hand, has the basic chromosome
number, x.
Constancy of genetic material through successive generation is essential for
maintaining the identity of species. But variation is essential for evolution to occur.
Such variation in chromosome number is called heteroploidy. Individuals carrying
chromosome numbers other than the diploid (2x, and not 2n) number are known as
heteroploids, and the condition is known as heteroploidy.
Numerical changes in chromosomes (heteroploidy) mainly of two types:
1. Euploidy and 2. Aneuploidy

Euploid
 Euploid derived from Greek words; Eu = true ; ploidy = unit
 The term euploidy designates a change in chromosome number which involves
entire set of chromosomes (x).
 Euploids have one or more number of complete chromosome sets (genomes),
which may be identical with or distinct from each other.
 The somatic chromosome number of a euploid individual is exact multiple of
basic chromosome number of that species.
 Euploidy includes monoploids, diploids and polyploids

Aneuploidy
 Variation in chromosome number which do not involve whole set of
chromosomes, but only a part of a set is referred as aneuploidy. It is any
deviation from a euploidy condition.
 This condition can be expressed either as an addition of one or more entire
chromosome or as a loss of such chromosomes as compared to the somatic
chromosome number of that species.
 Therefore, the Aneuploid is an organism or a cell having one or few
chromosomes more, or less than the normal somatic number (2n) of the
individual. It seems that the aneuploid changes in chromosome number do not
 involve the whole genome. They relate only one or few chromosomes of the
genome.

Origin of aneuploidy

1. Non-Disjunction
2. Loss of chromosomes
3. Irregularities of chromosome distribution

1. Non-disjunction: Generally during gametogenesis the homologous

chromosomes of each pair separate out (disjunction) and are equally
distributed in the daughter cells. The failure of separation of homologous
chromosome is called non-disjunction. It occurs during gametogensis. Here,
one type contains 22 chromosomes, while other will be 24.

2. Loss of chromosomes in mitotic or meiotic cells due to laggards (lagging

chromosomes), which are characterized by retarded movement during anaphase.
3. Irregularities of chromosome distribution during meiosis of polyploids with uneven
number of basic genomes like triploids and pentaploids.

Types of Aneuploids

Types Genomic constitution

Monosomic 2n – 1
Double monosomic 2n –1-1 Hypoploidy
Nullisomic 2n –2
Trisomic 2n +1
Double trisomic 2n +1+1 Hyperploidy
Tetrasomic 2n +2
Pentasomic 2n +3
1. Monosomy

The diploid organism which lacks one chromosome of a single homologous pair is
called monosomic with genomic formula 2n-1. A monosomic produces two types of
gametes n and n-1 because single chromosome without a pairing partner may go to
either of poles during meiosis.
The monosomics are usually weaker than normal diploids. Monosomics are normally
found in polyploids and the diploids cannot tolerate them.

The number of possible monosomics in an organism will be equal to the haploid

chromosome number. In common wheat, since 21 pairs of chromosomes are present,
21 possible monosomics are [Link] 21 monosomics in wheat were produced by
Sears in 1954 in the variety Chinese spring and are being used for genetic studies all
over the world. Monosomics have been used extensively in wheat breeding for the
purpose of chromosome substitution.

Double monosomics (2n-1-1) or triple monosomics (2n-1-1-1) could also be produced in

polyploids like wheat. In double monosomics the missing chromosomes are non-
homologous.

Genetic significance

 The monosomics are usually weaker than normal diploids.

 Monosomics are normally found in polyploids.
 Diploids cannot tolerate the loss results in high mortality or reduced
fertility.
 Useful in location of genes due to the presence of single copy of
chromosome.

Nullisomy

Diploid organisms which have lost a pair of homologous chromosomes are called
nullisomics with genomic formula 2n-2. In double monosomy and nullisomy, the
chromosome number is same but the genomic formula differs. In nullisomy (2n-2), a
complete homologous chromosome pair is missing. In double monosomy, (2n-1-1), two
chromosomes of different chromosome pairs (one each from two different
chromosome pairs) are missing.

 Double nullisomy (2n-2-2) involves loss of two pairs of homologous

chromosomes.
Nullisomics are not usually found in natural populations, but have to be obtained by
intercrossing or selfing of monosomics (2n-1). These can occur by fusion of two
gametes that are lacking in the same chromosome.

Genetic significance

 Nullisomic series are not of great agronomic importance, but used for genetic
studies.
 They exhibit reduced vigour, fertility and survival.
 Several nullisomics of hexaploid wheat (6x-2) exhibit reduced vigour and
fertility but can survive to maturity
 Produces distinct morphological effects – indicative of chromosomes involved.

Trisomy

Trisomics are those organisms which have one extra chromosome (2n+1). In plants,
the first case of trisomy was reported in Jimson weed, Datura stramonium (2n=24)
by Blakeslee and [Link] (1924). Since the extra chromosome may belong to any
one of the different chromosome pairs, the number of possible trisomics in an
organism will be equal to the haploid chromosome number. For instance, barley (2n =
14) the haploid chromosome number is n = 7. Consequently seven trisomics are
possible, in a trisomic, one of the pairs of chromosomes has an extra member and
forms a trivalent during anaphase I of meiosis. Two chromosomes will go to one pole
and one chromosome will go to other pole. As a result, two types of gametes are
formed i.e. n and n+1. This is very common in plants and has variable effects on
phenotype.

An individual having two extra chromosomes each belonging to a different

chromosome pair is called double trisomic (2n+1+1). Depending on the nature of extra
chromosome, simple trisomics are of three types.

a) Primary trisomics: The additional chromosome is normal one in primary trisomics.

b) Secondary trisomics: Trisomics having isochromosome as additional chromosome.
c) Tertiary trisomics: When additional chromosome in a trisomic is translocated one.

Meiotic behaviour
 During anaphase I of meiosis , one of the pairs of chromosomes has an extra
member and forms a trivalent.
 Two chromosomes will go to one pole and one chromosome will go to other
pole.
 As a result, two types of gametes are formed i.e. n and n+1. This is very
common in plants and has variable effects on phenotype.
 Selfing produces both normal and trisomic progenies

Genetic significance

 Trisomics are viable in diploid species and available in maize, tomato,

barley, bajra etc.
 Trisomy of different chromosome produces different morphological effects.
 In human being, trisomy 21 produces Down‟s syndrome called Mongolism.
 Trisomic analysis used for locating genes on specific chromosome. Ratio will
deviate in F2 from 3:1

Tetrasomy

Tetrasomics have a particular chromosome represented four times. Therefore the

general genomic formula for tetrasomics is 2n+2. When two chromosomes are added
each to two different pairs of chromosome (2n+2+2), it is double trisomic.
All the 21 possible tetrasomics in wheat are viable. Tetrasomics often behave more
regularly than the aneuploids with odd number of chromosomes.

Origin

Selfing of trisomy produces n+1 gamete. The union of n+1 gamete with another n+1
gamete produces tetrasomy

Meiotic behaviour

 The four homologues tend to form a quadrivalent , a trivalent & univalent

or two bivalents at meiosis.
 Quadrivalent disjoin as 2:2 or 3:1. Hence, tetrasomic plants produce n, n+1
and n+2 and zygotes with 2n, 2n+1 and 2n+2.

Aneuploidy condition in human beings

Autosomal aneuploidy
(i) Down‟s Syndrome:

It is frequent autosomal trisomic in human beings due to the trisomy for chromosome
21. First described by Langron Down of England. This syndrome is also known as
‗Mongolism‘ or ‗Mongolian idiocy‘. Some persons suffering from Down syndrome may
show the normal diploid chromosome number i.e. 46, instead of 47. But in these
persons, the long arm of chromosome no. 21 is found translocated onto another
chromosome of the complement.

Symptoms

Persons with Down‘s Syndrome shows a strong mental retardation. Their body is short
about 120 cm with stubby fingers. They possess wide nostrils, flattened nose, upward
slanting eyes, small ears, small mouth, swollen tongue, monkey-like skin ridges, short
hand and short fingers. The most prominent feature is the epicanthic fold—the
prominent eyelid folds like those of Mongolian people. Life expectancy is 8-16 yrs.

(ii) Edward‟s Syndrome:

It is due to the trisomic condition for the chromosome 18. The syndrome is
characterized by mental deficiency; multiple congenital malformations affect
virtually every organ system. Babies suffering from this syndrome usually die within a
year. In some rare cases they survive up to their teen years.
Sex chromosomal aneuploidy
(iii) Klinefelter‟s Syndrome (XXY)
It is due to the trisomic condition for the sex chromosomes (XXY). Individuals
possessing this syndrome are phenotypically males but with some tendency toward
femaleness, particularly in secondary sex characteristics. They show enlarged breasts,
less body hair, under developed testes and small prostrate glands. Naturally these
individuals remain sterile with retarded growth.
(iv) Turner syndrome (X0)

It is first discovered by H.H. Turner in 1938. Due to monosomy for X-chromosome

i.e. the individuals possess one normal X. Therefore adults with Turner Syndrome are
females having virtually no ovaries. The secondary sex characters are also poorly
developed. Besides, they show short stature, low set ears, webbed neck,a shield-like
chest, cardiovascular abnormalities and hearing impairment. But it is interesting
that these individuals generally do not show any mental retardation. It occur in one in
2500 live female birth, 90 % abort spontaneously

Applications of aneuploids

1) Aneuploids have been used to determine the phenotypic effects of loss or gain
of different chromosomes.
2) They are used to produce chromosome substitution lines (replacing
chromosome of one variety with chromosome of another variety of same
species). Such lines provide information on the effect of different
chromosomes of a variety in the same genetic background.
3) They are also used to produce alien addition (addition of chromosome from
related species) and alien substitution (chromosome(s) of one species
replaced by the chromosome(s) of another species).
4) Monosomics are also used in transferring chromosomes with desirable genes
from one species to another.
5) Aneuploid analysis permits the location of a gene as well as of a linkage group
on to a specific chromosome. Monosomics and nullisomics are used for this
purpose.
6) Studies on nullisomic and tetrasomic combinations made it possible to establish
homoeology among the chromosomes of A, B and D genomes of wheat.
7) Aneuploids are also useful in identifying the chromosomes involved in
translocations (tertiary trisomics).
8) Aneuploids are also useful in the preparation of molecular maps.
Lec 10: Polyploid - auto and allopolyploids, their characters; meaning of genome;
evolution of wheat, Triticale, cotton, tobacco, Brassica
Polyploids: An individual having more than two basis sets of chromosomes.
Autopolyploids: Polyploids which originate by the multiplication of chromosomes of a
single species.
Autotriploids: An autopolyplod having three set of chromosomes of the same species.
Autotetraploids: An autopolyplod having four copies of genome of same species.
Allopolyploids: A polyploid which combines complete genome from two or more
species.
Amphidiploid: An allopolyploid which arises by combining the genomes of two
diploid species, behaves as a diploid during meiosis.
A segmental allopolyploid contains two or more genomes, which are identical with
each other, except for some minor differences.
Allosyndesis: In allopolyploid, the pairing between the genomes of two different
species(Intergenome pairing).
Autosyndesis: Intra-genomal pairing in allopolyploids.
Colchiploidy:Colchicine induced polyploidy.
Disomic: Normal diploid species.
Diploid: An individual with two sets of basic chromosome number.
Diploidization: The process by which a polyploidy species behaves like a diploid
species.
Homologous pairing: The pairing between homologues chromosome of the same
genome (Intra-genome pairing)
Homeologous pairing: The pairing between the genomes of two different species
(Intergenome pairing).

Euploids have one or more complete genomes, which may be identical with or
distinct from each other. The somatic chromosome number of a euploid individual is
exact multiple of basic chromosome number of that species. Euploidy includes
monoploids, diploids and polyploids.

Monoploids & Haploids

Monoploids denote the presence of a single copy of a single genome or basic
chromosome number (x) whereas haploids representing the gametic chromosome
number of a species (n).
Polyploidy

This means, organisms showing polyploidy possess more than two sets of chromosomes
in their nuclei. It may be 3 (tripoid), 4 (tetraploid), 5 (pentaploid), 6 (hexaploid), 7
(heptaploid), 8 (octaploid) or more genomes making up their somatic chromosome
number.

However, beside monoploids and polyploids, another category known as diploids is

found. But diploids do not represent any deviation. Rather, they convey the normal
condition of the organisms. The diploid individuals possess two sets of homologous
chromosomes – one paternal and one maternal.

Occurrence of Euploids

1. Most animal species are diploid. Polyploidy in animals is generally lethal.

2. Some naturally occurring euploidy variations eg. bees - females are diploid;
drones are monoploid (ie haploid).
3. Some amphibian & fish polyploids are known.
4. Plants commonly exhibit polyploidy. About 30-35% of ferns and flowering plants
are polyploidy. Many of the fruits & grain are polyploid plants.

Monoploids & Haploids

Monoploids: Monoploids contain a single chromosome set and are characteristically

sterile. In other words, monoploids have the basic chromosome number (x) of a
species.
Haploid: Haploid is a general term used to designate the individuals or tissues with a
gametic chromosome number i.e. n. Haploids (n) carry half or gametic chromosome
number.
In a true diploid species, both monoploid and haploid chromosome number are same
(i. e. x=n).

 Example: Maize 2n = 20 ; x = 10 & n = 10

 Hexaploid wheat Wheat 2n = 6x = 42; x = 7 & n = 21

Haploids are of two types

1. Monohaploids: Individuals which arise from a normal diploid species.

Eg. : Maize 2n = 20 and n = 10
2. Polyhaploids: Individuals which arise from any polyploid species.
Eg : Hexaploid Wheat 2n = 6x = 42 and n=3x=21
Origin of Haploids
Haploids can arise spontaneously or can be induced. Spontaneous haploids have been
obtained in plants like tomato, cotton, barley, pearl millet and wheat. The first
induced haploid was produced by Jorgensen in 1928 by crossing Solanum nigrum x
Solanum luteum. Guha and Maheswari (1964) obtained haploid plants of Datura
innoxia directly from the pollen by culturing the anthers. The main reason to obtain
the haploids has been to develop a new and rapid method of breeding homozygous
diploids or polyploids through diploidization using colchicine. Diploids obtained
through the chromosome doubling of haploids are known as dihaploid.

Characteristic features of haploid plants

 Haploids are smaller, less vigorous than their diploid phenotypes.
 Haploids are sterile, as the chromosomes have no regular pairing partner
homologous chromosomes during meiosis and they are found as univalents at
metaphase I of meiosis.
 The meiotic products are deficient in one or more chromosomes.

Polyploidy
The organisms with more than two genomes are called polyploids.

Generation of Polyploids
1. Complete non-disjunction of both gametes can produce a polyploidy individual.
2. The creation of autotriploids can be accomplished by crossing a tetraploid with a
Diploid.
3. Interspecific crosses can generate Allopolyploids.
4. They can be produced through chromosome doubling of a species using colchinine.
Colchicine is an alkaloid extracted from the seeds of Colchicum autumnale.
Colchicine interferes with the development of spindle apparatus, as a result sister
chromatids are unable to move to opposite poles during [Link]
retained in the same nucleus (restitution) leading to chromosome doubling

Autopolyploids: When all the genomes present in a polyploidy species are identical, it
is known as autopolyploid and the situation in termed as autopolyploidy. It includes
triploids (3x), tetraploids (4x), pentaploids (5x), hexaploids (6x), septaploid (7x),
octoploids (8x) and so on.

Autotriploids : Three set of chromosomes of same species. They can occur naturally
or can be produced by crossing an antotetraploid and diploid species. They are highly
sterile due to defective gamete [Link] are useful which are propagated
asexually.
Autotriploids:They have four copies of geneome of same [Link] may arise
spontaneously or induced by doubling the chromosome of a diploid species with
colchicines treatment. They are stable and fertile because pairing partners are
available during meiosis.
Genetic significance of autopoplyploids
 Most autoployploid species show increase in vigour and size knows as gigantism.
Leaves are larger and thicker, flowers, fruits and seeds are also larger.
 Cell size, pollen grains and stomata are relatively larger.
 Grwoth rate are generally lower and their flowering is later than normal
diploids.
 Increase in fresh weight due to more water content.
 It has multivalent formation leading to unbalanced gamete formation.

Application of autopolyploidy in crop improvement

1. Triploids :
Triploid water melon: Seedless water melons are produced by crossing
tetraploids (4x), used as female with diploids (2x) as male. The reciprocal cross
is not successful.
Triploid sugar beet : Have larger roots and more sugar per unit area than
diploids and tetraploids. 3x is the optimum level of ploidy in sugar beets. Seed
production is done by planting 4x and 2x plants.
Triploid banana: Once triploids are produced they are propagated
vegetatively.

2. Auto Tetraploids :
Grapes: Developed in California which has large fruits and fewer seed than
diploids.
Rye : Grown in Sweden and Germany which have large seeds and high protein
content
Alfalfa :Better in yield and recovery after grazing
Berseem : Pusa Giant variety is cultivated for forage purpose.
Ornamental : Tetraploids are successful, because of increased flower size and
longer flowering period.

Limitations
1. Autopolyploids are successful in species with lower chromosome number.
2. Cross pollinated species are more responsive than self pollinating species.
3. Larger size of autopolyploids is generally accompanied with higher water
content. This is not a desirable character in cabbage and turnip.
4. In crops grown for seed exhibit high amount of sterility though seed size is
increased.
 5. New autopolyploids cannot be used directly as crops because they will have
some undesirable traits e.g. poor strength of stem in grapes, irregular fruit size
in water melon.
6. Effects of autopolyploidy cannot be predicted.

Allopolyploids:
A polyploid which originates by combining complete genome from two or more
species. It is also known as hybrid polyploids or bispecies or multispecies
polyploids. They are developed by interspecific crosses and fertility restoration by
colchicine treatment. It played a major role in evolution & 50% plants are
allopolyploids. Based on the origin, classified into
1. Natural allopolyploids and [Link] allopolyploids

Natural allopolyploids
1. Origin of Hexaploid wheat

Natural Allopolyploids
Origin of hexaploid wheat
[Link] x T. speltoides
AA BB
2n = 14 2n = 14
F1 Sterile (2n=14) (AB)
Natural Doubling

[Link] 2n = 28 x [Link] (Aegilops squarrosa)

AABB DD
2n = 28 2n = 14
ABD (2n= 21) Sterile
Natural doubling
[Link]
AABBDD (2n = 42)
(Cultivated)
 2. Evolution of tetraploid cotton
2a. Evolution of tetraploid cotton
2b. Evolution of tetraploid cotton
Origin was postulated by Beasley (1940)
Origin was postulated Philips (1963)
G. arboreum x G. thurberi
G. herbaceum x G. raimondii
AA DD
AA DD
2n = 26 2n = 26
2n = 26 2n = 26
AD AD
2n = 26 (Natural Chromosome 2n = 26
doubling)
(Natural Chromosome doubling)
AA DD
2n = 52 AA DD
G. hirsutum (tetraploid cotton) 2n = 52
G. barbadense (tetraploid cotton)

3. Evolution of cultivated tobacco

[Link] of cultivated tobacco

Nicotiana sylvestris x N. tomentosa

2n = 24 2n = 24

F1(2n=24)

(Chromosome doubling)

N. tabacum
Cultivated tobacco
(2n=48)

4. Evolution of Brassica species

4. Evolution of Brassica species (Origin was given by Nigraha U)

[Link] (Black
n=8 mustard)
BB

(Abyssinian [Link] [Link] (Indian mustard)

cabbage) n=17 n=18
BBCC AABB

[Link] [Link] [Link]

n=9 n=19 n=10
CC AACC AA
(Cabbage) (Rapeseed) (Rapeseed)

U triangle Rape seed and Mustard

Artificial allopolyploid
1. Raphanabrassica :
It was synthesized by Russian Scientist – Karpechenko in 1927 with an objective to
develop a fertile hybrid with the root of radish and leaves of cabbage. The species
used were Radish-Raphanus sativus (2n=18) and Cabbage-Brassica oleracea (2n =18).

However, fertile amphidiploid (2n=36) derived by spontaneous chromosome doubling

had the root of cabbage and leaves of radish and hence of no use.

2. Evolution of Triticale- Man made cereal

Triticale was evolved by crossing tetraploid wheat (2n = 28 -AABB) or hexploid

wheat [Link] (2n=6x=42-AABBDD) with Rye (Secale cereale 2n = 14 RR
genome)
Role of Allopolyploids and their evolution
1. About 1/3 of angiosperms are polyploids. These suggest that polyploids have
significant role in the evolution of crop species.
2. Allopolyploids have contributed great extent in the evolution of plants than
auto polyploids.
3. The identification of diploid parental species is primarily based on pairing
between the chromosome of diploid and the allotetraploid species.
4. Allopolyploids combine the genome of different species, hence the resulting
individuals differ from progenitor.
5. Evolution is a slow process; but due to allopolyploids new species originate
very quickly.
6. Polyploids have wider adaptation to different environmental condition.

Terms to describe different types of euploidy

Lec 11: Pre-Mendelian ideas about heredity – Vapour and fluid theory, Magnetic
power theory, Preformation theory, Lamarck‟s theory, Darwin‟s theory,
Germplasm theory and Mutation theory

[Link] and fluid theory

Moist vapour theory: Early Greek Philosopher Pythagoras (580-500 BC) proposed the
theory that a moist vapour descends from the brain, nerves and other body parts
of the male during coitus and from this, similar parts of embryo are formed in the
uterus of the female.
Fluid theory: Empedocles (504-433 BC) proposed the theory that each parent
produces semen which arises directly from various parts of the body. In embryos the
various parts are formed by this semen.

2. Magnetic Power Theory

During 17th Century, William Harvey (1578-1657) after performing certain experiments
on deer proposed the theory called Magnetic Power Theory. He suggested that as iron
by friction with a magnet possesses the magnetic properties. Like that, the uterus by
the friction of coitus acquires some magnetic power to conceive an embryo.

3. Preformation Theory
The theory was proposed by two Dutch biologists, Swammerdam and Bonnet (1720-
1793). This theory states that a miniature human called „homunculus‟ was already
preformed in the egg and sperm. The development of zygote resulted only in the
growth of miniature human, who was already present in the egg and sperm.
However, this theory was rejected because this could not be proved scientifically.
4. Theory of Epigenesis
This theory was proposed by Wolf (1738-1794), a German biologist. This theory states
that egg or sperm cells do not contain miniature human but that the gametes
contained undifferentiated living substance capable of forming the organized body
after fertilization. This concept is known as epigenesis, which is universally
accepted.
5. Theory of Acquired characters
This concept was proposed by Lamarck (1744-1829), a French biologist. This theory
states that a new character once acquired by an individual shall pass on to its
progeny. This theory was disproved by Weismann. He cut the tail of mice for
successive generations and always got the baby mice with tail. Thus, this theory was
rejected.

6. Theory of Pangenes
This theory was proposed by Charles Darwin (1809-1882), an English naturalist.
According to him, each part of the animal body produces a minute copy of its own,
called gemmule or pangene. The gemmules are collected in the reproductive organs.
The gemmulues were then given to the gametes. The young one formed from the
gametes would be having all the gemmules characteristics of the parents, and will
represent a blending of the qualities of its two parents. Thus, theory of pangenesis is
a theory of „blending inheritance‟

7. Evolutionary theory of inheritance by natural selection

It was proposed by Charles Darwin. He believed that evolution is due to natural
selection of small hereditary variations occurring among individuals of any species.
Darwin defined evolution as "descent with modification" the idea that species change
over time, give rise to new species, and share a common ancestor. The mechanism
that Darwin proposed for evolution is natural selection. Because resources are limited
in nature, organisms with heritable traits that favor survival and reproduction will
tend to leave more offspring. He published his findings in the book ―Origin of Species‖

8. Germplasam Theory
This theory was advocated by August Weismann (1834-1914), a German biologist.
According to this theory, organism‘s body contains two types of cells namely somatic
cells and reproductive cells. The somatic cells form the body and its various organ
systems, while the reproductive cells form sperm and ova. The somatic cells contain
the „somatoplasm‘ and reproductive cells contain the ‗germplasm‟. The germplasm
can form somatoplasm, but somatoplasm cannot form germplasm.
The cells of the somatoplasm become differentiated during the formation of the
complex organs of the body, while cells of the germ cells remain undifferentiated and
retain their power to generate new life. The germplasm thus goes on in continuous
stream from generation to generation. Changes in the somatic cells (somatoplasm),
which were caused by the environment, cannot influence the germplasm and hence
acquired characters are not inherited.

9. Mutation theory of inheritance

The theory was proposed by Hugo De Vries in 1901. He has introduced the term
mutation for these large discontinuous variations in the genotype and proposed the
mutation theory of inheritance. According to this, sudden hereditary changes lead to
evolution. He observed that some plants of Oenothera lamarckiana differ in some
characters from the typical O. lamarckiana. It is a self fertilized crop the variations
were accounted due to sudden changes rather than hybrids. He found that these
variations were inherited and called these changes as mutations and play a role in the
evolution of new species.
Lec 12 : Mendel‟s experiments and laws of inheritance. Rediscovery of Mendel‟s
work.
Definitions
Genes: A basic unit of inheritance/heredity. A sequence of DNA nucleotides which
coded for a functional product of RNA or a ploypeptide. The factors which are
responsible for the inheritance of characters from one generation to another.
Allele : An allele is an alternate form of a gene, which are located at the same
position or genetic locus, on a chromosome
Contrasting characters: Features of an individual with marked (observable)
phenotypic difference.
Phenotype: The observable characteristics of an individual resulting from the
interaction of its genotype with the environment (or) the actual appearance of an
individual. [Link], height etc.,
Genotype: The genetic constitution of an individual such as RR, Rr or rr.
F1:The first filial generation or offspring of two distinct parents.
F2: The progeny of F1 plants obtained by selfing.
Dominance: Masking effect of one allele of a gene over the other.
Dominant: The character which is expressed in F1.
Recessive: The character whose effect is suppressed or masked in F1.
Gene symbol: Various symbols which are used to represent genes or alleles.
Homozygous: Individuals having identical alleles on the corresponding locus of
homologous chromosomes.
Heterozygous: Individuals having dissimilar alleles on the corresponding locus of
homologous chromosomes.
Locus: A position on chromosome which is occupied by an allele.
Hybridization / crossing: The process through which the hybrids are produced
involving parents having contrasting characters. Hybridization cross may be a
monohybrid cross, dihybrid cross or polyhybrid cross depending upon the kind of
hybrid it produces.
Hybrid : Product of a hybridization is called hybrid.
Monohybrid cross: Crossing of the two individuals differing in one pair of gene
affecting one character.
Dihybrid cross: Crossing of the two individuals differing in two pairs of gene affecting
two characters.
Polyhybrid cross: Crossing of the two individuals differing in three or more pairs of
genes, each affecting a different character.
Direct cross: A cross between two genotypes.
Backcross: Crossing F1 with any one of the parents.
Testcross: Crossing F1 with homozygous recessive parent.
Pre- mendelian work on plant hybridization
A number of scientists worked on plant hybridization during 18 th and 19th centuries.
Kolreuter (1760) was the first scientist involved in plant hybridization experiments.
He observed uniformity and heterosis in F1 and variation in F2.
Gaertner (1722-1820) has carried experiments on Garden pea.
Knight, Seton and Goss: studied experimenting with inheritance of seed colour in
garden pea.
Naudin (1815-1909) and Darwin (1809-1882) also hybridized plants and got similar
results and confirmed the observations of Kolreuter. However, they could not give
an explanation for their results.

Reason for failure of Mendel‟s earlier workers

1. Studied large number of characters at a time
2. Characters not classified into clear cut classes
3. Frequency of different characters forms in the progeny were not made properly
4. The data from different generations were not kept accurate and separately
5. Complete control of pollination not enforced
6. Interspecific hybrids exhibited partial and complete sterility
7. F2 population was small
8. Most of the characters are quantitative nature

Gregor Johann Mendel and his work

Gregor Johann Mendel was a Monk Born in 1822 in Silesia near Austria (now Czech
Republic). He is a son of peasant farmer, studied Theology and was designed priest.
In his youth, he led a disastrous, poor, difficult and sad life. In the year 1846, Johann
Gregor Mendel attended courses of agriculture, pomiculture and viniculture at the
Philosophical Academy in Brunn.
He went to the University of Vienna and studied botany. In 1856 Mendel began
experiments on plant hybridization with purelines of garden pea (Pisum sativum) in
Monastery garden for eight years. He has cultivated and tested some 29,000 pea
plants between 1856 to 1863. He was the first person to work out the basic laws that
governs the inheritance of genes. Gregor Johann Mendel has first offered necessary
explanation for the inheritance and hence he is known as ―Father of Genetics‖.

From these experiments he deduced two generalizations which later became known
as Mendel's Laws of Heredity or Laws of inheritance. He described these laws in a
paper, "Experiments on Plant Hybridization" presented to the Natural History
Society of Brunn on February 8 and March 8, 1865. In 1866, his paper ―Experiments
on plant hybridization‖ published in volume 4 of the proceedings of the Natural
Science Society. In the same year, he began experiments with other plant species. In
this paper, Mendel proposed some basic genetic principles. But unfortunately his
remarkable piece of work remained unattended and unappreciated upto 1900.

Mendel‟s selection of the Experimental Plant

Mendel chosen garden pea (Pisum sativum) as the plant material, since it has the
following advantages.
1. Convenience of handling: Each plant occupies only a small space grown easily
either in field or in pots pace.
2. Controlled mating: The flower structure of pea ensures self pollination, which was
experimentally verified by Mendel. Flowers are relatively large, emasculation and
pollination is quite easy. Therefore, crossing could be carried out easily.
3. Short life cycle: Peas complete their life cycle from seed to seed within 70 days,
thus producing many generations in rapid succession.
4. Large number of fertile off-springs:
Hybrids resulted from crossing two pure strains (true breeding) were perfectly
fertile and more in number. Seeds are large in size and do not have any problem in
germination.
5. Presence of variation:
Peas have many sharply defined inherited differences like plant height (Tall vs.
dwarf), seed shape (round vs wrinkled) etc. In the available varieties, several
characters had two contrasting forms, which were easily distinguishable from each
other. This permitted an easy classification of F2 and F3 progenies.

Mendel‟s Experimental Observations and Results

1. Mendel‘s observations of the hybrids (F1) revealed that hybrid plants contained
the character of only one parent, none of them displayed any intermediate
character of both parents.
2. Those characters which were transmitted unchanged and expressed in the
hybrid are called as dominant characters and those which became latent in
the process are called as recessive characters.
3. He tested the all seven pairs of contrasting characters for the phenomenon of
the dominance and recessiveness and found following characters as dominant.
4. Mendel‘s observations of F2 progeny further revealed the fact that the recessive
character which was depressed or concealed in F1 hybrids reappeared in the F2
offspring in the definitely expressed average proportion of three to one, so that
among each four plants of F2, three displayed the dominant character and one
the recessive character.
5. This 3:1 F2 ratio was observed to occur in the monohybrid cross for each of
seven pairs of characters.
Reasons for success of Mendel‟s work
1. The experiments were very well designed and conducted with great care and
skill.
2. The choice of experimental material, the common garden pea.
3. Mendel studied the inheritance of only one pair of contrasting characters at a
time.
4. The characters he chose were well defined and simple; each with only two
contrasting forms Eg: Seed coat colour of peas is either green or yellow, with
no intermediate types.
5. The seven characters selected by Mendel showed qualitative inheritance.
6. The contrasting forms of each of the seven characters were governed by a
single gene and in each case one form was completely dominant over other.
7. His greatest innovation was to count the number of progeny in each category to
emerge from a given cross for every generation.
8. His knowledge on mathematics was a definite asset for the interpretation of his
findings after subjecting the results to more refined mathematical analysis.
9. He maintained particulars of pedigree records, which gave him the exact
ancestry of any given plant.
Reason for the negligence
i. Mendel used mathematics to explain the principles of inheritance (New to the
biologists)
ii. The information on cell and cell division were insufficient
iii. Dalton supported Darwin‘s theory – continuous variation and its significance in
evolution. In Mendel‘s work no new form of variation appeared only parental
forms come into light- this is opposite to the theory of evolution. Did not fit
with scheme of biological evolution
iv. Mendel was later busy with his administrative work as Abbot of the Monastry .

Mendel‟s laws
Mendel himself did not postulate any genetical principle or laws. He simply gave
conclusive theoretical and statistical explanations for his hybridization experiments in
his research paper.
It was Correns the discoverer of Mendel‘s work who thought that Mendel‘s discovery
could be represented by the two laws of heredity. These laws of heredity are
1. Law of Segregation
2. The Law of Independent Assortment

The law of segregation or the law of purity of gametes

 It states that ―when a pair of alleles is brought together in a hybrid (F 1) they
remain together without contaminating each other and they separate or
 segregate from each other into a gamete in a complete and pure form during
the formation of gametes‖.
 The law is universal in its application and it has been found to occur in plants
as well as animals.

Mechanism of gene transmission

Explanation of Law of Segregation

Law of segregation or purity of gametes can be explained by considering the
monohybrid ratio - inheritance of only one character. In pea, round seed shape is
dominant over wrinkled seed shape. Two different alleles of the same gene i.e. ‗R‘
and ‗r‘ were brought together in the hybrid (F1). Even though the hybrid was round
seeded in the next generation (F2) it produced both round and wrinkled seeded
progeny. In F2 generation, the different phenotypes could be recovered because the
two alleles in F1 remained pure and did not contaminate each other thus producing
two types of gametes with (R) and (r). The separation of homologous chromosomes
during anaphase I of meiosis may be regarded as the reason for segregation of the
two alleles of a gene. This is because the alleles of a gene are located in an identical
position in the two homologous chromosomes.
 An individual possesses two
alleles for each trait; one
allele is given by the female
parent and the other by the
male parent
 When gametes form, the
paired alleles separate
randomly so that each
gamete receives a copy of
one of the two alleles.
 In heterozygous individuals
the only allele that is
expressed is the dominant.
The recessive allele is present
but its expression is hidden.
Mendel‟s Law of Independent Assortment
 The Law of Independent Assortment, also known as "Inheritance Law"
 The law states that “segregation of one pair of alleles is independent from
the segregation in any other pair of alleles during gamete formation”

Explanation of Law of Independent Assortment

i. Law of independent assortment can be explained by studying the inheritance of

two characters at a time, simultaneously (dihybrid cross).
ii. Independent segregation for two genes can be explained by assuming that the
two genes are located in two different chromosomes. The two alleles of a
gene will be located in the two homologues of the concerned chromosome.
iii. Independent separation of these two pairs of chromosomes at anaphase I of
meiosis will lead to the independent segregation of the genes located in them.
Thus any allele of one gene is equally likely to combine with any allele of
the other gene and pass into the same gamete.
iv. Independent segregation of two genes produces four different types of gametes
in equal proportion. A random union among these gametes gives rise to sixteen
possible zygotes. These zygotes yield a [Link] phenotypic ratio, which is
known as the typical dihybrid ratio.
v. When two pairs of independent alleles enter into F 1 combination, both of them
have their independent dominant effect. These alleles segregate when gametes
are formed. But the assortment occurs independently at random and quite
freely.

Mendel's experiments with

mixing one trait always
resulted in a 3:1 ratio
between dominant and
recessive phenotypes, his
experiments with mixing two
traits (dihybrid cross) showed
[Link] ratios.

The [Link] table shows that

each of the two genes are
independently inherited with
a 3:1 ratio
Rediscovery of Mendel's work
Mendel‘s research paper remained dormant and unnoticed by the scientific world
until 1900. It was in the beginning of 20th century that three botanists, namely Hugo
de Vries, working on Oenothera ; Correns working on Xenia, peas and maize and
Von Tschermak working on various flowering plants, independently drawn the
conclusions like Mendel. Later these botanists came across the research paper of
Mendel and rediscovered it in 1900. Mendel‘s original paper was republished in
Flora, 89, 364 (1901). Bateson confirmed Mendel‘s work by a series of hybridization
experiments.

Backcross : Crossing of a hybrid with one of its parents.

Test cross : Crossing of a hybrid with

recessive parent.

To determine whether an individual with

a dominant phenotype is homozygous for
the dominant allele or heterozygous,
Mendel crossed the individual in question
with an individual that had the recessive
phenotype
Lec 13- Terminologies: gene, allele, locus, homozygous, heterozygous,
hemizygous, genotype, phenotype, monohybrid, dihybrid, trihybrid, polyhybrid.

Gene : It is an unit of heredity which is transferred from a parent to offspring and is

held to determine some characteristics of the offspring. A sequence of DNA
nucleotides which codes for a functional product of RNA or a ploypeptide.
Allele : An allele is an alternate form of a gene, which are located at the same
position or genetic locus, on a chromosome
Locus: A locus (plural loci) in genetics is the position on a chromosome. Each
chromosome carries many genes. Each gene occupies a specific locus on the
chromosomes
Homozygous: It refers to individuals having identical alleles for a single trait. [Link],
rr
Heterozygous: It refers to individuals having two different alleles for a specific gene.
[Link]
Hemizygous: Having only single copy of a gene instead of two copies. All the genes
on the single X chromosome in the male and Y chromosome are hemizygous.
Genotype: The genetic constitution of an individual organism
Phenotype: The set of observable characteristics of an individual resulting from the
interaction of its genotype with the environment
Monohybrid: A hybrid that is heterozygous for the alleles of one gene.
A monohybrid cross is a mating between two individuals with different alleles at one
genetic locus of interested gene
Dihybrid: A hybrid that is heterozygous for alleles of two different genes.
Dihybrid cross is a cross between two different lines (varieties, strains) that differ in
two observed traits
Polyhybrid: A hybrid whose parents differ in a number of characters.

Punnett Square
 Developed by Reginald Punnett.
 The Punnett square is a square diagram that is used to predict the genotypes of
a particular cross.
 The diagram is used by biologists to determine the probability of an offspring
having a particular genotype.
 Lec 14- Chromosomal theory of inheritance. Allelic interactions – Dominance vs
recessive, complete dominance, co-dominance, incomplete dominance, over
dominance, threshold characters.
Dominant : The allele which can phenotypically express itself in heterozygote and
homozygote is called as dominant trait.
Recessive : The allele which can phenotypically express only in homozygote is
called as recessive trait.

Chromosomal theory of inheritance

In 1902, Walter Sutton, Theodor Boveri have proposed chromosome theory of
inheritance, also known as Sutton-Boveri theory. It is the fundamental theory of
genetics which identifies chromosomes as the carriers of genetic material. The
Principles of segregation and independent assortment are explained by the
behaviour of chromosomes during meiosis.

The theory states that

– Mendelian factors (genes) have specific loci on the chromosomes
– Chromosomes undergo segregation and independent assortment.

Evidence that Genes Associated to Chromosomes

Thomas Hunt Morgan was the first to associate a specific gene with a specific
chromosome. He has provided convincing evidence that chromosomes are the
location of Mendel‘s heritable factors.
Experimental insect- Drosophila melanogaster, a fruit fly species that eats fungi on
fruit.
– Fruit flies are prolific breeders with generation time of two weeks.
– Fruit flies has three pairs of autosomes and a pair of sex chromosomes
(XX in females, XY in males).

Wild Type and Mutant Phenotypes

Wild type Red eyes
After a year of research, Morgan discovered a mutant /variant (traits alternative to
the wild type) male fly with White eyes
In one experiment Morgan mated female flies with red eyes (wild type) with male
flies with white eyes (mutant)
– The F1 generation all had red eyes
– The F2 generation showed the 3:1 red:white eye ratio
The white-eyed trait appeared only in males. All the females and half the males had
red eyes
Morgan concluded that a fly‘s eye color was linked to its sex.
Morgan‟s Discovery
The transmission of the X chromosome in fruit flies correlates with inheritance of
the eye-color trait. This was the first solid evidence indicating that a specific gene is
associated with a specific chromosome

Types of allelic interaction (intra-allelic interaction)

1. Complete dominance 2. Incomplete dominance
3. Co-dominance 4. Over dominance

1. Complete dominance

One allele is completely

Monohybrid Cross: Complete Dominance
dominant to another. Example
Phenotype of the heterozygote Cross two heterozygous red (Rr) F1 offspring.
is the same as homozygous What is the genotype and phenotype of their F2 offspring?
1. Parental genotypes = Rr x Rr
dominant. 2. Assign gametes
3. Assign offspring Eggs
Recessive phenotype is genotypes
4. Assign phenotypes Sperm R (.5) r (.5)
expressed only when the 5. Results:
1 RR = 25%
organism is homozygous 2 Rr = 50% 100% R (.5) Rr (.25)
RR (.25)
recessive. 1 rr = 25%
6. Genotypic Ratio = [Link] Homozygous Dominant Heterozygous
Red Flower
Red Flower

e.g., Mendel‘s garden pea 7. Phenotypic Ratio = 3:1

traits r (.5) Rr (.25) rr (.25)

Heterozygous Homozygous Recessive
Red Flower White Flower
Exceptions of Mendel‟‟s findings
1. Incomplete dominance
When a dominant allele does not
mask completely the phenotypic
expression of the recessive allele in a
heterozygote, then a blending of
both dominant and recessive traits
takes place in the F1 and F2
heterozygotes. This phenomenon is
known as incomplete or partial
dominance.

The F1 gametes produce F2 progeny

having the phenotypic and genotypic
ratios of 1 : 2 : 1. (Example: A cross
between Red & White flowered
Mirabilis jalaba)

2. Co-dominance
Both dominant and recessive
alleles lack their dominant and
recessive relationships and both
have capability to express them
phenotypically in the heterozygous
condition. In a heterozygote of co-
dominant nature, the dominant and
recessive traits occur side by side.
The F1 heterozygotes produce a F2
progeny in the phenotypic and
genotypic ratios of 1 : 2 : 1 like the
incomplete dominance. (Example:
Coat Colour in Cattles)

Human blood type

A group=IAIA, IAi ; B group=IBIB, Ibi
AB group =IAIB ;O group= ii
Alleles IA for A antigen is co-dominant with the allele IB for B antigen. The dominant
relation symbolized as (IA = IB) >i

The main difference between codominance and incomplete dominance lies in the way
in which genes act. In case of codominance both alleles are active while in case of
incomplete dominance both alleles blend to make an intermediate one.

3. Overdominance/hetero/super dominance
When the heterozygotes have a more extreme phenotype than either of the
corresponding homozygotes (homozygous parents), then it is referred to as
overdominance / superdominance / heterodominance.
For example, when the heterozygote Aa between a pair of factors which control size
is bigger than the homozygotes AA or aa. (Eg : Hybrids in Plants).

Threshold characters
 A threshold trait is a trait, which is inherited quantitatively, but is expressed
qualitatively.
 Involves the relationship between polygenes and discontinuous variation.
 Those polygeneically determined genotypes have values below threshold show
no expression of the character.
 Expression only when the genotypes have values above this threshold.

Two scales are involved

i. Underlying Polygenic distribution, which is continuous
ii. Superficial phenotypic distribution, which is discontinuous

Relationship between two scales was demonstrated by Wright in cross between

strains of guinea pigs.
i. One strain-normal three toes in hind leg & other was polydactylous with four
toes.F1-three toed ; F2-3:1 ratio of three toed to four toed.
iii. In test cross (F1 x recessive parent)-77 %-four toed & 23 % three toed.
iv. He proposed that 4 pairs of polygenes or eight alleles involved in polydactyly.
v. Individuals with five or more polydactylous alleles out of eight exceeded the
threshold & appeared as four toed.
Lec 15- Deviation from Mendelian inheritance – Non allelic interaction without
modification in Mendelian ratio – Bateson and Punnett‟s experiment on fowl comb
shape. Non allelic interaction with modification in Mendelian ratio – i.) Dominant
epistasis ([Link])

Definitions
Epistasis: Interaction of two or more genes, thus involving two or more loci
Dominant Epistasis: Gene interaction in which a dominant allele at one locus can
mask the expression of both alleles (dominant and recessive) at another locus
resulting in 12 : 3 : 1 ratio . Also referred as simple epistasis.

TYPES OF GENE ACTION

The interaction within the alleles of gene controlling a single character may be
dominant, incomplete dominance and co-dominance and are called intra genic
interaction. When there is an interaction occurs between different pairs of alleles
influencing a character of an individual is said to be intergenic or epistatic
interaction.
Gene interaction or epistasis
The term epistasis was coined by Bateson in 1909. In epistatic interactions,
different genes located on the same or different chromosomes interact with one
another for the expression of a single phenotypic trait of an organism. When two
different genes interact and affect the same character, such a genetic interaction is
said to be intergenic or non-allelic. Gene interaction or epistasis is the functional
interaction between different genes that occur when an allele at one locus mask or
inhibit the expression of a non-allele at another locus. Any gene that masks the
expression of another non-allelic gene is called epistatic, while the gene that is
masked or suppressed is hypostatic.

[Link] Dominance Epistasis

1. Interaction between two alleles Interaction between alleles of two or

of the single gene more genes

2. Involves only single locus Involves two or more loci

3. It is known as intragenic or It is known as intergenic or non-allelic

inter-allelic gene interaction gene interaction

4. Dominance is of three types viz., Epistasis is of several types viz.,

complete, incomplete and co- dominant epistasis, recessive epistasis,
dominance duplicate dominant, duplicate
recessive
5. Partial dominance alters the It modifies the normal dihybrid ratio in
normal segregation ratio of 3:1 F2
into [Link]

6. Recessive genes can express only Recessive genes can also exhibit
in homozygous condition masking effect

Gene interaction without modification of

Mendelian ratio (9 : 3 : 3 : 1)

Two pairs of alleles affecting the same character and producing in the F 2 four
different phenotypes in the ratio of 9 : 3 : 3 : 1 was discovered in fowls by Bateson
and Punnett.
Each breed of poultry possesses a characteristic type of comb. Each of these breeds
true.

– The Wyandotte breed has ‗Rose‘ comb

– The Brahma has a ‗Pea‘ comb,
– The Leghorn has a ‗Single‘ comb
– Malay breed has a comb - ‗Walnut‘ comb.
Cross between rose comb and single combed types show that rose in dominant to
single comb and that there is a segregation of 3 rose: 1 single comb in the F 2. In
mating between pea combed with single combed and 3:1 ratio appears in F 2. In
mating between pea combed with single combed bird, pea combed is found to be
dominant over single comb and 3:1 ratio appears in F2.

When a rose combed fowl is crossed with a pea combed one, all the F 1 birds showed a
new comb known as walnut comb. When the walnut combs are inbred there appears
in F2 walnut, rose pea and single comb in the ratio of [Link]. The rose comb is due
to the presence of R gene and Pea due to P gene. Walnut comb is due to the presence
of the dominant genes. R and P and single comb are due to the presence of recessive
of r and p. The ratio expected in F2 is [Link].

Parents RR PP x rrpp
Rose x pea

Rr pp(Walnut)
 ♀/♂ RP Rp rP rp
RRPP RRPp RrPP RrPp
RP
(W) (W) (W) (W)
RRPp RRpp RrPp Rrpp
Rp
(W) (R) (W) (R)
RrPP RrPp rrPP rrpp
Rp
(W) (W) (P) (P)
RrPp Rrpp rrPp rrPP
Rp
(W) (R) (P) (S)
9 Walnut:3 Rose: 3 Pea : 1 Single

Types of epistasis
Epistasis leads to modification of normal dihybrid or trihybrid segregation ratio
in F2 generation. Various types of epistatic gene interaction are

1) Dominant epistasis ([Link])

2) Recessive epitasis ([Link])
3) Duplicate genes with cumulative effect ([Link]).
4) Duplicate dominant epistasis (15:1)
5) Duplicate recessive epistasis (9:7) and
6) Dominant and recessive epistasis (13:3)
1. Dominant Epistasis ([Link])

Gene interaction in which a dominant allele at one locus masks the expression of both
alleles (dominant and recessive) at another locus resulting in 12 : 3 : 1 ratio. It is also
referred as simple epistasis.
An example of dominant epistasis is found for fruit colour viz., white, yellow and
green. White colour is controlled by dominant gene W and yellow colour by dominant
genes G. White is codominant over both yellow and green. The green fruits are
produced in recessive condition (wwgg). A cross between plants having white and
yellow fruits produced F1 with white fruits. Intermating of F1 plants produced plants
with white, yellow and green coloured fruits in F2 was [Link] ratio. Here W is
dominant to w and epistatic to alleles G and g. Hence it will mask the expression of G
and g alleles. Hence in F2 plants with W-G- (9:16) and W-gg (3:16) genotypes will
produce white fruits; plants with wwG-3/16 will produce yellow fruits and those with
wwgg 1/16 genotype will produce green fruits. Thus the normal dihybrid ration
[Link] is modified to [Link] ratio in 1:2 generation. Similar type of gene interaction
has been reported for skin colour in mice and seed coat colour in barley.

Parents White fruit x Yellow fruit

WWgg x wwGG

WwGg White fruit

♂
♀ WG Wg wG Wg

WWGG WWGg WwGG WwGG

Wg
(W) (W) (W) (W)
WWGg WWgg WwGg Wwgg
Wg
(W) (W) (W) (W)
WwGG WwGg wwGG WwGg
WG
(W) (W) (Y) (Y)
WwGg Wwgg wwGg wwgg
Wg
(W) (W) (Y) (G)
Ratio = 12 White: 3 Yellow: 1 Green
Lec 16 & 18- ii.) Recessive epistasis ([Link]) iii.) Duplicate and additive epistasis
([Link]). iv.) Duplicate dominant epistasis (15:1); v) Duplicate recessive epistasis
(9:7) vi.) Dominant and recessive epistasis (13:3); Summary of epistatic ratios (i)
to (vi).
Definitions

Recessive epistasis: Gene interaction in which recessive allele at one locus masks the
expression of both the dominant and recessive alleles at other locus resulting in
9 : 3 : 4 ratio. It is also called as supplementary gene action.
Duplicate genes with cumulative effect: Gene interaction in which two dominant
alleles have similar effects when they are separate, but produce enhanced effect
when come together, resulting in 9 : 6 : 1 ratio in F2. Also called as additive gene
action.
Duplicate dominant epistasis: Gene interaction in which a dominant allele at either
of the loci can mask the expression of recessive alleles at the two loci, resulting in
15 : 1 ratio ; also referred to as duplicate gene interaction.
Duplicate recessive epistasis: Gene interaction in which recessive alleles at either of
two loci can mask the expression of dominant alleles at the two loci, resulting in 9 : 7
ratio ; also called as complementary gene action.
Dominant and Recessive epistasis: The dominant allele at one locus (I-) and recessive
alleles at second locus (pp) produces same phenotype resulting in 13 : 3 ratio. Locus P-
expressed only when I is in reccessive condition (aa); Also known as inhibitory gene
action.

[Link] epistasis ( [Link]) (Supplementary gene action)

Gene interaction in which recessive allele at one locus can mask the expression
of both the alleles at other locus resulting in 9 : 3 : 4 ratio. Also called as
supplementary epistasis.
Here one dominant gene has its own phenotypic effect and other dominant gene
has no effect of its own but its presence with the first gene modified the phenotypic
expression. Thus in supplementary gene action, the dominant allele of one gene is
necessary for the development of the concerned phenotype, while the other gene
modifies the expression of the first gene.

[Link] colour in Maize

Three colors - Purple, Red and White

• Purple color - presence of 2 dominant genes (R and P)
• Red color -presence of 1 dominant gene (R)
• White color – presence of recessive gene rr(rrP- and rrpp)
 Parents RR PP X rr pp
Purple Red
RP rp
RrPp
Purple
♂
RP Rp RP Rp
♀
RP RRPP RRPp RrPP RrPP
(P) (P) (P) (P)
Rp RRPp rrPP RrPp rrPp
(P) (W) (P) (W)
Rp RrPp RrPp RRpp Rrpp
(P) (P) (R) (R)
rp RrPp rrPp Rrpp rrpp
(P) (W) (R) (W)
Ratio = 9 Purple : 3 Red : 4 White
• The allele r is recessive to R but epistatic to alleles P and p.
• In F2, plants with genotypes
R-P- (9/16) – Purple color grains
R-pp (3/16) – Red color grains
rrP- (3/16)) – White color grains
rrpp (1/16) – White color grains
The dihybrid [Link] ratio is modified to [Link] ratio
The epistatic allele is r – produce white color under rrP- homozygous recessive
condition.
P-Hypostatic gene

[Link] genes with cumulative effect ([Link]) (Additive gene action)

Gene interaction in which two dominant alleles have similar effects when they
are separate, but produce enhanced effect when come together, resulting in 9 : 6 : 1
ratio in F2.
In these two genes controlling a character produces identical phenotype when
they are alone i.e. with the homozygous recessive condition of the other gene. But
when both the genes are present together, their phenotype effect is enhanced as if
the effect of the two genes were cumulative or additives. It should be noted that in
this case both the genes show complete dominance. In barley two completely
dominant genes A and B affect the length of awns, the thin needle like extension of
lemma genes A and B alone (e.g. Aabb and aaBB give gives rise to awn of medium
length, the effect of A is the same as that of B. But when both the genes A and B are
present together they produce long awn indicating the effect of A and B genes of awn
length are added together. Individual homozygous recessive for both these genes are
awn less.

Parents AA BB x aa bb
Long awned x awnless

AaBb
Long Awned
♂
AB Ab aB ab
♀
AABB AABb
AaBB AaBB
AB (L) (L)
(L) (L)
AABb
Ab AAbb AaBb Aabb
(L)
(A) (L) (A)

aB AaBB AaBb aaBB aaBb

(L) (L) (A) (A)
AaBb Aabb aaBb aabb
ab
(L) (A) (A) (a)

Ratio = 9 Long awned: 6 Awned: 1 awnless

Another example: Fruit shape in Summer Squash

Three types of fruit shape

 Disc - 2 dominant genes A and B

 Spherical – either gene A or gene B
 Elongated – double recessive aabb

A cross between
Disc X Elongated
AABB aabb
F1 Disc shaped AaBb

Intermating of F1
D
In F2, Plants with genotypes
A-B- (9/16) – Disc shaped fruits
A-bb (3/16) and aaB- (3/16) – Spherical fruits
aabb (1/16) – Elongated fruits
F2 AB Ab aB ab

AB AABB AABb AaBB AaBb

Disc Disc Disc Disc
Ab AABb AAbb AaBb Aabb
Disc Spherical Disc Spherical

aB AaBB AaBb aaBB aaBb

Disc Disc Spherical Spherical

ab AaBb Aabb aaBb aabb

Disc Spherical Spherical Elongated

4. Duplicate dominant epistasis (15:1) (Duplicate gene action)

Gene interaction in which a dominant allele at either of the loci can mask the
expression of recessive alleles at the two loci, resulting in 15 : 1 ratio ; also referred
to as duplicate gene interaction.
Eg: Awn character in Rice & Pericarp colour in rice

In rice awn character is controlled by two dominant duplicate genes (A and B).
Presence of any of these two alleles can produce awn. The awnless condition develops
only when both these genes are in homozygous recessive state (aabb). A cross
between awned and awnless strains produced awned plants in F1. Intermating of F1
plants produced awned and awnless plants in 15:1 ratio in F2 generation. The allele A
is epistatic to a/b alleles and all plants having allele A will develop awn. Another
dominant allele B is epistatic to alleles a/b. An individual with these allele also
develop awn character.

awned awnless
Parents rice x rice
AAbb x aaBB

AaBb
Awned
rice
 ♂
♀ AB Ab aB ab

AB AABB AABb AaBB AaBb

(A) (A) (A) (A)
Ab AABb AAbb AaBb Aabb
(A) (A) (A) (A)
aB AaBB AaBb aaBB aaBb
(A) (A) (A) (A)
ab AABb AAbb AaBb Aabb
(A) (A) (A) (a)

Ratio = 15 awned : 1 awnless

In F2, plants with genotypes

A-B- (9/16), A-bb (3/16) and aaB- (3/16) – develop -AWN
aabb (1/16) double recessive condition – develop- AWNLESS

5. Duplicate recessive epistasis (Complementary gene action- 9:7)

Gene interaction in which recessive alleles at either of two loci can mask the
expression of dominant alleles at the two loci, resulting in 9 : 7 ratio ; also called
complementary gene action. When recessive alleles at either of the two loci can mask
the expression of dominant alleles at the two loci, it is called duplicate recessive
epistasis. This is also known as complementary epistasis.
The best example of duplicate recessive epistasis if found for flower colour in sweet
pea. The purple colour of flower in sweet pea is governed by two dominant gene say A
and B when these genes are in separate individuals (Aabb or aaBB) and white (aabb)
they produce white flower. A cross between purple flower (AABB) and white flower
(aabb) strains produced purple colour in F1/. Intermating of F1 plants produced purple
and white flower plants in 9:7 ratio in F2 generation. Here the recessive allele ‗a‘ is
epistatic to B/b alleles and mask the expression of these alleles, another recessive
allele b is epistatic to A/a alleles and mask their expression.

Parents purple x White

AABB x aabb
AB ab
AaBb
Purple
Selfing of F1

9 Purple flower
and 7 white flower
in F2
 ♂
AB Ab aB ab
♀

AB AABB AABb AaBB AaBb

(P) (P) (P) (P)

Ab AABb AAbb AaBb Aabb

(P) (W) (P) (W)

aB AaBB AaBb aaBB aaBb

(P) (P) (W) (W)

Ab AaBb Aabb aaBb aabb

(P) (W) (W) (W)

Ratio = 9 Purple : 7 white

 In F2, plants with genotypes
o A-B- (9/16)– develop PURPLE flowers
o A-bb (3/16), aaB- (3/16) and aabb (1/16) – develop WHITE flowers

6. Dominant and Recessive epistasis (13:3) (Inhibitory gene action)

The dominant allele at one locus (I-) and recessive alleles at second locus
(pp) produces same phenotype. Locus P- expressed only when I is in recessive
condition (aa) ; Also known as inhibitory gene interaction.

An example of this type of gene interaction is found for anthocyanin pigmentation in

rice. The green colour of plants is governed by the gene I which is dominant over
purple colour. The purple colour is controlled by a dominant gene P. When a cross
made between green (IIpp) and (iiPP) colour plants, the F 1 was green. Intermating of
F1 plants produced green and purple plants in 13:3 ratio in F2.

• Green color of plants is governed by gene “I”

• Purple color is governed by the dominant gene “P”
• The gene “I” is dominant over gene “P”
A cross was made between
Green X Purple
IIpp iiPP
F1 Green IiPp
Intermating of F1 -13 green and 3 purple plants (13:3) in F2
The allele “I” is epistatic to alleles P and p
In F2, plants with genotypes
I-P- (9/16), I-pp (3/16), and iipp (1/16) – were GREEN
―I” will mask the effect of P or p
iiP- (3/16) – were PURPLE (“I” is absent)

♂
IP Ip iP ip
♀
IP IIPP (G) IIPp (G) IiPP (G) IiPP (G)

Ip IIPp (G) IIpp (G) IiPp (G) Iipp (G)

iP IiPP (G) IiPp (G) IiPP (P) iiPp (P)

ip IiPp (G) Iipp (G) iiPp (P) Iipp (G)

13 green : 3 purple plants

Lec 16- Lethal genes, Pleiotrophy, penetrance and expressivity, phenocopy: Multiple
alleles, blood group in humans, coat colour in rabbits, self incompatibility in plants; pseudo
alleles, isoalleles.
Definitions

Lethal alleles: are alleles that cause the death of the organism that carries them. They are usually
a result of mutations in genes that are essential to growth or development. Lethal alleles may be
recessive or dominant depending on the genes involved.
Pleiotropism: A phenomenon in which a gene has more than one phenotypic effects. Such genes
are called pleiotropic genes.
Penetrance: The frequency with which a gene produces a phenotypic or visible effect in the
individuals, which carry it. Two types are complete and incomplete.
Complete penetrance: Expression of a gene in all the individuals which carry it.
Incomplete penetrance: Expression of a gene in less than 100 % of its carriers.
Expressivity: The degree of phenotypic expression of a penetrant gene in its carriers. It is of two
types uniform and variable.
Uniform Expressivity: Similar or uniform expression of a gene in all the individuals that carry
such gene.
Variable Expressivity: Differentiation or variable expression of a gene in the individuals that
carry it.
Phenocopy: The alteration of the phenotype, by nutritional factors or the exposure to
environmental stress during development to form imitating phenotype that characteristically
produced by a gene. Eg. Rickets due to lack of vitamin D deficiency would be a phenocopy of
vitamin D resistant rickets.
Multiple alleles: Existence of more than two alleles at a locus.
Lethal gene: Gene which causes death of its carrier when in homozygous condition.
Self incompatibility: The inability of the fertile pollen grains to fertilize the same flower.
Pseudo alleles: Closely linked and functionally related genes. A cluster of pseudo alleles is
known as complex locus or complex region. Eg. Lozenge and star asteroid eyes in Drosophila.
Isoalleles: An allele which is similar in its phenotypic expression to that of other independently
occurring alleles.
Transgressive segregation
The appearance in F2 individuals with higher or lower intensity of characters than the parents is
called as transgressive segregation.
Atavism, throw back or reversion: The reappearance of ancestoral characters after several
generations because of recessiveness or other masking effect.
Essential Genes and Lethal Genes
Presence of lethal allele will results in the death of an organism. Gene that is involved in the
death is called the essential gene. Two types.
– Dominant lethal allele
– Recessive lethal allele
Recessive lethal allele
1. Yellow body color in mice

Yellow (AY) is dominant over non-

yellow (A)
– AY/ AY = lethal
– AY/ A = yellow
– A/ AY= non-yellow
2:1 ratio of hetero yellow and non-
yellow

2. Recessive lethal in human beings

i. Tay-Sachs – enzyme deficiency that prevents nerve function
ii. X-linked lethal genes- Hemophilia
Dominant lethal genes : very rare
i. Manx gene in cats – Genotype- Mm: tail-less; mm: normal ; MM: dead
ii. Huntington’s Disease -H : type of neurotoxin h : no neurotoxin
iii. Retinoblastoma: R : allows tumor formation r : no tumor formation
iv. Sickle-cell disease (SCD), or sickle-cell anaemia (or anemia; SCA) or
drepanocytosis, is an autosomal co-dominant genetic blood disorder characterized
by red blood cells that assume an abnormal, rigid, sickle shape. Sickling decreases
the cells' flexibility and results in a risk of various complications.
Pleiotropy : A phenomenon in which one gene affecting more than one phenotypic characters.
Such genes are called pleiotropic genes.
i. E.g.,Waardenburg syndrome - White forelock, pale iris, deafness
ii. Sickle cell allele

Penetrance
The frequency with which a gene produces a phenotypic or visible effect in the individuals,
which carry it, is known as penetrance. In other words penetrance refers to the proportion of
individuals which exhibit phenotypic effect of a specific gene carried by them. In general, genes
express themselves in all the individuals in which they are present in the appropriate genotype is
known as penetrance. It indicates the number of individuals that give the expected phenotype to
any degree.
1. Complete Penetrance
i. Most dominant and recessive genes in homozygous conditions and many completely
dominant genes even in heterozygous condition give their complete phenotypic
expressions. Such genes which always produce the expected phenotype are having
complete penetrance.
ii. If only 70 per cent of the individuals of a stock homozygous for a certain recessive gene
show the character phenotypically, the gene is said to have 70 per cent penetrance.
Examples of Complete Penetrance
i. In pea, the alleles (RR) for red flowers and the alleles (rr) for white flowers have
complete penetrance in homozygous conditions.
ii. In Drosophila the recessive alleles for vestigial wings in homozygous conditions have
complete penetrance.
iii. In guinea pigs the dominant allele ‘B’ for black coat has complete penetrance both in
homozygous and heterozygous conditions.
2. Incomplete Penetrance
 Some genes in homozygous as well as in heterozygous conditions fail to provide
complete (cent per cent) phenotypic expression of them. Such genes are called to have
incomplete penetrance.
Examples of Incomplete Penetrance
i. Polydactyly is a condition with extra fingers and toe or toes in man is due to the presence
of dominant gene P. The normal condition is produced by the genotype PP. The genotype
and pp produce polydactyly. Some heterozygous individual are not polydactly (Pp).
Therefore the gene has penetrance of less than 100 per cent and said to be incompletely
penetrant.
ii. In, man the tendency to develop Diabetes mellitus (a condition in which there is an
excess of sugar in the blood) is controlled by certain genes. However, not everyone
carrying the genes for diabetes actually develops the conditions, hence these genes have
an incomplete penetrance.
Expressivity
The degree of phenotypic expression of a penetrant gene is called expressivity. In other words,
the ability of a gene to produce identical phenotypes in all the individuals carrying it in the
appropriate genotype is known as complete expressivity.
Examples of Expressivity
Polydactyly
The degree of expression produced by a penetrant genotype is termed expressivity. The
polydactylous condition may be penetrant in the left hand and not in the right hand or may be
penetrant in the feet and not in hands.
Phenocopy: is a variation in phenotype (generally referring to a single trait) which is caused by
environmental [Link] environmentally induced change which resembles the effect of
gene mutation. It can last only for that generation in which the environment that induces the
change is present. The term proposed by Richard Goldschmidth.
Eg. Temperature- Some enzymes are affected or catalyzed by temperature. Function at one
temperature but not by the other. In Himalayan rabbits, cold temperature causes dark fur to
develop.
Multiple alleles: More than two allelic forms of a gene occupy the same locus of the
homologous chromosome
1. Multiple Alleles & Codominance in humanbeings
 i. There are four phenotypes: A, B,
AB, and O
ii. The ABO blood group is controlled
by three alleles viz.,IA, IB, and IO
iii. Each individual inherits only two
alleles for these genes.
iv. IA, IB are codominant, IA and IB
have dominance over i

A person has two alleles. They can be:

i. two A antigen (blood type A)
ii. two B antigen (blood type B)
iii. one each of A and B antigens (blood type AB)
iv. No antigen (blood type O)
Human blood types can be
type A (IAIA or IA i)
type B (IBIB or IBi)
type AB (IAIB)
type O (ii)

ABO blood group compatibility

Blood group Can donate to: Can receive from:

A A or AB A or O

B B or AB B or O
AB (universal recipient) AB only A, B, AB, or O

O (universal donor) A, B, AB, or O O only

ABO Transfusion Compatibility

i. Both the IA and IB alleles put a protein on the RBC’s that is recognized by the immune
system.
ii. The immune system of a Type-A person will attack any RBC’s with B protein (cannot
accept B or AB blood).
iii. The immune system of a Type-B person will attack any RBC’s with A protein (cannot
accept A or AB blood).
iv. AB person will accept both A and B proteins.
v. The IO allele does not put any protein on the rbc’s, therefore, type O blood is accepted by
anyone.
vi. The immune system of a Type-O person will attack any rbc’s with either A or B protein
(can only accept type O)

2. Fur colour in Rabbits

The fur colour is of four types – Agouti, Chinchilla, Himalayan and Albino

Agouti

• This has full colour, known as wild type

• This colour is dominant over all the remaining colours
• Produces agouti colour in F1 and 3:1 in F2 when crossed with any of the other three
colours in rabbits.
• This colour is represented by C+
• C+ C+ , C+ Cch , C+ Ch , C+ c
Chinchilla
• This is lighter than Agouti.
• This colour is dominant over Himalayan and albino
• Produces chinchilla in F1 and 3:1 ratio in F2 when crossed with either Himalayan or
albino.
• This colour is represented by cch
• Genotypes cch cch cch c
Himalayan
• The main body is white while the tips of ear, feet and tail and snout are coloured
• This colour is dominant over albino
• Produces 3:1 ratio in F2 when crossed with albino.
• This is represented by Ch Ch Ch c
Albino

• This has pure white fur colour and is recessive to all other types.
• This is represented by c
Phenotypes and genotypes of multiple allelic series
 PHENOTYPES GENOTYPES

Agouti C+C+, C+Cch, C+Ch, C+c

Chinchilla CchCch,

Light grey CchCh, Cchc

Himalayan ChCh, Chc

Albino cc
3. Self imcompatibility system in plants: The inability of the fertile pollen grains to
fertilize the same flower. Controlled by a series of multiple alleles- S1 > S2 > S3 >S4

4. Seed coat pattern in Lentil

5 different alleles - spotted, dotted, clear (no pattern), marble-1, marble-2

Characteristics features of the multiple alleles

i. The alleles occupy the same locus.
ii. Only two members of such alleles are present at a time in a diploid organism
iii. There is no crossing over in the multiple allelic series(since they are present in the same
locus)
iv. Wild type is always dominant. Others exhibit dominant or intermediate phenotypic
expression.
v. Two mutant alleles will produce mutant phenotype – never produce wild type i.e. no
complementation
 vi. It controls the same character of an individual

Pseudo alleles : Pseudo alleles refer to closely linked and functionally related genes.
Characteristics of pseudo alleles
i. Pseudo alleles govern different expressions of the same character
ii. Pseudo alleles occupy a complex locus, which is divided into sub loci
iii. They exhibit low frequency of genetic recombination by crossing over
iv. They exhibit cis-trans position effect.

Iso alleles : An allele that is similar in its phenotypic expression to that of other independently
occurring allele is known as isoallele. Isoalleles are two types.
i. Mutant isoalleles : such alleles act within the phenotypic range of a mutant character
ii. Normal isoalleles : such alleles act within the phenotypic range of a wild character

Polygenic inheritance : Additive effects of two or more genes on a character

Eg. Skin pigmentation (atleast 3 genes)

Hair color (atleast 4 genes)
 PBG 201 Principles of Genetics and Cytogenetics (2+1)
Theory Notes (After midterm)

Lec 19- Lethal genes, Pleiotrophy, penetrance and expressivity, phenocopy: Multiple
alleles, blood group in humans, coat colour in rabbits, self incompatibility in plants; pseudo
alleles, isoalleles.

Lethality : The phenotypic manifestation of some genes lead to the death of the individual in
prenatal or postnatal prior to maturity.
Lethal genes: are genes that cause the death of the organism that carries them. They are usually
a result of mutations in genes that are essential to growth or development. Lethal alleles may be
recessive or dominant depending on the genes involved.
Dominant lethal gene : A gene which kills the individual both in homozygous and heterozygous
condition arises by mutation from normal allele.
Recessive lethal allele: Genes which cause death of the individual in homozygous condition.
Pleiotropy : The phenomenon of multiple phenotypic expressions of a single gene i.e a gene
influences more than one trait.
Pleiotropic gene: A gene that influences more than one trait.
Penetrance: The frequency with which a gene produces a phenotypic effect in the individuals,
which carry it. Most dominant and recessive genes in homozygous conditions and many
dominant genes in heterozygous condition give their complete phenotypic expressions.
Incomplete penetrance: Some genes in homozygous as well as in heterozygous conditions fail
to provide complete (100 per cent) phenotypic expression of them i.e. expression of a gene in
less than 100 % of its carriers.
Expressivity: The degree of effect produced by a penetrant genotype is called expressivity.
Expressivity is a variation in the strength of a [Link] is of two types uniform and variable.
Uniform Expressivity: Similar or uniform expression of a gene in all the individuals that carry
such gene.
Variable Expressivity: Differentiation or variable expression of a gene in the individuals that
carry it.
Phenocopy: It is the variation in phenotype is not due to genotype, but due to environmental
conditions.
Multiple alleles: Presence of more than two alleles of a gene occupy the same locus on the
homologous chromosome controlling a particular trait is termed as multiple alleles.
Self incompatibility: The inability of the fertile pollen grains to fertilize the ovary of the same
flower.
Pseudo alleles: Pseudo alleles refer to alleles of two or more tightly linked genes, affecting the
same function. A cluster of pseudo alleles is known as complex loci.
Isoalleles: An allele which is similar in its phenotypic expression to that of other independently
occurring alleles.
Lethal gene: Gene which causes death of its carrier.
[Link] lethal gene
A gene which kills the individual both in homozygous and heterozygous condition arises by
mutation from normal allele. Individuals with dominant lethal die before they produce progeny.
Hence the dominant lethal mutant removed from the population in same generation.
[Link] lethal gene
Genes which cause death of the individual in homozygous condition . Lethal genes discovered
when severe distortion of Mendelian ratios in the progenies.

Eg . [Link] body color in mice

Yellow (AY) is dominant over

non-yellow (A)
– AY/ AY = lethal
– AY/ A = yellow
– A/ AY= non-yellow
2:1 ratio of hetero yellow and non-
yellow

2. Recessive lethal in human beings

Sickle cell anaemia

Homozygous recessive HbsHbs die prematurely
Ratio of 1 HbAHbA :2 HbAHbs
Change of monohybrid cross ratio to 2:1 instead of [Link]

Dominant lethal genes : very rare

i. Manx gene in cats – Genotype- Tt tail-less; tt: normal ; TT: lethal
ii. Huntington‟s Disease -H : type of neurotoxin h : no neurotoxin
iii. Retinoblastoma: R : allows tumor formation ;r : no tumor formation
 3. Sublethal or Subvital or semilethal genes
A recessive lethal gene that lacks complete penetrance and expressivity will kill most, but not
all homozygotes before sexual [Link] lowers viability.

4. Balanced lethal system : First discovered by Muller in Drosophila for Curly wing.
Wing shape – Dominant gene Cy; Eye colour – Dominant gene Pm. Both genes are
located on the same chromosome and homozygous. They causes lethality both in
dominant/recessive condition.

Cy+Pm cypm

Cy+Pm Cy+Pm /Cy+Pm Cy+Pm /cypm

Lethal Curly plum

cypm Cy+Pm / cypm cypm /cypm

Curly plum Lethal

Pleiotropy - The phenomenon of multiple phenotypic expressions of a single gene. Such genes
are called pleiotropic genes.
In Drosophila, the recessive gene for vestigial wings causes vestigial wings in homozygous
condition. In addition to this, following traits are also affected :
(i) the tiny wig like balancer behind the wings
(ii) certain bristles
(iii) the structure of the reproductive organs
(iv) egg production is lowered and longevity is reduced

Pleiotropism in humanbeing
[Link] syndrome (PKU)
In human, the gene for disease phenylketonuria has pleiotorpic effect and produces
various abnormal phentoypic traits, collectively called syndrome. It also causes secretion
of excessive quantity of amino acid phenylalanine in urine, cerebrospinal fluid and
blood, short stature, mental retardation, widely spaced incisors and pigmented patches on
skin, excessive sweating and non-pigmented hairs and eyes.
2. Sickle-cell anemia
Sickle-cell disease (SCD), or sickle-cell anaemia (SCA) or drepanocytosis, is an
autosomal co-dominant genetic blood disorder characterized by red blood cells that
assume an abnormal, rigid, sickle shape. It is due expression of a single mutated HBB
gene produces numerous consequences throughout the body. The mutated hemoglobin
forms polymers and clumps together causing the deoxygenated red blood cells to assume
 the sickle shape. These cells are inflexible and cannot easily flow through blood vessels,
increasing the risk of blood clots and possibly depriving vital organs of oxygen. Some
complications associated with sickle cell anemia include pain, damaged organs, strokes,
high blood pressure and loss of vision. Sickle red blood cells also have a shortened
lifespan and die prematurely.

3. Waardenburg syndrome - White forelock, pale iris, deafness

Penetrance
The frequency with which a gene produces a phenotypic in the individuals, which carry it, is
known as penetrance. In other words penetrance refers to the proportion of individuals which
exhibit phenotypic effect of a specific gene carried by them. In general, genes express
themselves in all the individuals in which they are present in the appropriate genotype is known
as penetrance. It indicates the number of individuals that give the expected phenotype to any
degree.
1. Complete Penetrance
Most dominant and recessive genes in homozygous conditions and many completely
dominant genes even in heterozygous condition give their complete phenotypic
expressions. Such genes which always produce the expected phenotype are having
complete penetrance.
Examples of Complete Penetrance
i. In pea, the alleles (RR) for red flowers and the alleles (rr) for white flowers have
complete penetrance in homozygous conditions.
ii. In Drosophila the recessive alleles for vestigial wings in homozygous conditions have
complete penetrance.
 iii. In guinea pigs the dominant allele „B‟ for black coat has complete penetrance both in
homozygous and heterozygous conditions.
2. Incomplete Penetrance
Some genes in homozygous as well as in heterozygous conditions fail to provide
complete (cent per cent) phenotypic expression of them. Such genes are called to have
incomplete penetrance.
Examples of Incomplete Penetrance
i. Polydactyly is a condition with extra fingers and toe or toes in man is due to the presence
of dominant gene P. The normal condition is produced by the genotype PP. The genotype
and pp produce polydactyly. Some heterozygous individual are not polydactly (Pp).
Therefore the gene has penetrance of less than 100 per cent and said to be incompletely
penetrant. In one study, 65% of people with the dominant allele were born with extra
fingers and/or toes. The rest had normal fingers and toes. Thus polydactyly has 65%
penetrance (the percentage that expressed the trait).
ii. In, man the tendency to develop Diabetes mellitus (a condition in which there is an
excess of sugar in the blood) is controlled by certain genes. However, not everyone
carrying the genes for diabetes actually develops the conditions, hence these genes have
an incomplete penetrance.
Expressivity
The degree of phenotypic expression of a penetrant gene is called expressivity. In other
words, the ability of a gene to produce identical phenotypes in all the individuals carrying it in
the appropriate genotype is known as complete expressivity.
Examples of Expressivity - Polydactyly
The degree of expression produced by a penetrant genotype is termed expressivity. The
polydactylous condition may be penetrant in the left hand and not in the right hand or may be
penetrant in the feet and not in hands.

Phenocopy
It is the variation in phenotype is not due to genotype, but due to environmental
[Link] environment change the expression of a gene and the resulting phenotype is
the same that produced by another allele of the same gene. This type of effect due to
environment is phenocopy effect and the individuals are phenocopiesis a variation in phenotype
(generally referring to a single trait) which is caused by environmental conditions. It can last
only for that generation in which the environment that induces the change is present. The term
proposed by Richard Goldschmidth.
Eg. 1. Drosophila-Body colour- Y
Y— grey body colour
yy- yellow body colour
If larvae with yy genotype cultured in nutrient medium rich in silver salt, produces phenotype of
dominant allele.
2. Some enzymes are catalyzed by temperature. Function at one temperature but not in the other.
In Himalayan rabbits, cold temperature causes dark fur to develop.

Multiple alleles: More than two allelic forms of a gene occupy the same locus of the
homologous chromosome controlling a particular trait.
1. Multiple Alleles & Codominance in humanbeings

i. There are four phenotypes: A, B,

AB, and O
ii. The ABO blood group is
controlled by three alleles viz.,IA,
IB, and IO
iii. Each individual inherits only two
alleles for these genes.
iv. IA, IB are codominant, IA and IB
have dominance over i

Any person can have only two alleles. It may be

i. two A antigen (blood type A)
ii. two B antigen (blood type B)
iii. one each of A and B antigens (blood type AB)
iv. No antigen (blood type O)
Human blood types can be
type A (IAIA or IA i)
type B (IBIB or IBi)
type AB (IAIB)
type O (ii)
• Alleles I for A antigen is co-dominant with the allele IB for B antigen.
A

• The dominant relation symbolized as (IA = IB) >i

ABO blood group compatibility

Blood group Can donate to Can receive from

A A or AB A or O

B B or AB B or O

AB (universal recipient) AB only A, B, AB, or O

O (universal donor) A, B, AB, or O O only

ABO Transfusion Compatibility

i. Both the IA and IB alleles put a protein on the RBC‟s that is recognized by the immune
system.
ii. The immune system of a Type-A person will attack any RBC‟s with B antigen (cannot
accept B or AB blood).
iii. The immune system of a Type-B person will attack any RBC‟s with A antigen (cannot
accept A or AB blood).
iv. AB person will accept both A and B antigens.
v. The IO allele does not put any protein on the rbc‟s, therefore, type O blood is accepted by
anyone.
vi. The immune system of a Type-O person will attack any rbc‟s with either A or B protein
(can only accept type O)

2. Fur colour in Rabbits

The fur colour is of four types – Agouti, Chinchilla, Himalayan and Albino

Agouti

• This has full colour, known as wild type

• This colour is dominant over all the remaining colours
 • Produces agouti colour in F1 and 3:1 in F2 when crossed with any of the other three
colours in rabbits.
• This colour is represented by C+ and genotypes C+ C+ , C+ Cch , C+ Ch , C+ c
Chinchilla
• This is lighter than Agouti and dominant over Himalayan and albino
• Produces chinchilla in F1 and 3:1 ratio in F2 when crossed with either Himalayan or
albino.
• This colour is represented by Cch and Genotypes Cch Cch Cch c
Himalayan
• The main body is white while the tips of ear, feet and tail and snout are coloured
• This colour is dominant over albino and roduces 3:1 ratio in F2 when crossed with albino.
• This is represented by Ch Ch Ch c
Albino
• This has pure white fur colour and is recessive to all other types.
• This is represented by c

PHENOTYPES GENOTYPES
Agouti C+C+, C+Cch, C+Ch, C+c
Chinchilla CchCch,
Light grey CchCh, Cchc
Himalayan ChCh, Chc
Albino cc

3. Self imcompatibility system in plants: The inability of the fertile pollen grains to fertilize
the same flower and controlled by a series of multiple alleles- S1 > S2 > S3 >S4
Two types : 1. Sporophytic system and 2. Gametophytic system

1. Sporophytic system - SI is governed by the genotype of the plant

Genotypes of
S1 S2 S1 S3 S1 S4 S2 S3 S2 S4 S3 S4
the plants
Genotype of the
(S1) (S2) (S1) (S3) (S1) (S4) (S2) (S3) (S2) (S4) (S3) (S4)
gametes
Incompatability
reaction of the All S1 All S1 All S1 All S2 All S2 All S3
pollen grain
Incompatability
reaction of the S1 S1 S1 S2 S2 S3
style
 2. Gametophytic system-SI is governed by genotype of the gametes

[Link] coat pattern in Lentil

Five different alleles determines five traits viz., spotted, dotted, clear (no pattern), marble-
1, marble-2

5. Mice: coat colour

Agouti gene: (wild-type AA) overall appearance = grey and tip of hair black and yellow
atat-black back white belly ; aa-black
Dominance series : AA> atat> aa

Characteristics features of the multiple alleles

i. The alleles occupy the same locus.
ii. controls the same character of an individual
iii. Only two members of such alleles are present at a time in a diploid organism
iv. There is no crossing over in the multiple allelic series(since they are present in the same
locus)
v. Wild type is always dominant. Others exhibit dominant or intermediate phenotypic
expression.
vi. Two mutant alleles will produce mutant phenotype – never produce wild type i.e. no
complementation

Pseudo alleles : Pseudo alleles refer to alleles of two or more tightly linked genes, affecting the
same function.
Characteristics of pseudo alleles
i. Pseudo alleles govern different expressions of the same character
ii. Pseudo alleles occupy a complex locus, which is divided into sub loci
iii. They exhibit low frequency of genetic recombination by crossing over
iv. They exhibit cis-trans position effect.
 Eg. Lozenge locus i.e eye with glossy smooth surface in Drosophila and mapped at one
locus
Mutant – heterozygote with two different mutant genes produces lozenge phenotype
Iz1+/Iz2+ - Trans-heterozygote (mutant phenotype)
Iz1Iz2 /++ - Cis-heterozygote (wild phenotype)

Iso alleles : An allele that is similar in its phenotypic expression to that of other independently
occurring allele is known as isoallele.
Eg. Two alleles : +1 and +2 –indistinguishable
Isoalleles are two types.
i. Mutant isoalleles : such alleles act within the phenotypic range of a mutant character
ii. Normal isoalleles : such alleles act within the phenotypic range of a wild character
Lecture 20 &21- Quantitative inheritance – Multiple factor hypothesis – Nilsson Ehle
experiment on wheat kernel colour. Polygenes – transgressive segregation, comparison of
quantitatively and qualitatively inherited characters; modifiers;

Quantitative inheritance: The inheritance of many different genes influencing the same
character in a cumulative fashion is called as quantitative inheritance / multiple factor inheritance
/ polygenic inheritance.
Quantitative characters: Quantitative characters are controlled by poly genes (many genes)
with cumulative effect without dominance and produces continuous variation.
Polygene or multiple factor or quantitative gene: They are member of a group of non-
epistatic genes that interact additively to influence a quantitative trait.
Oligogenes or major genes: These genes have larger effect on the characters and shows
discontinuous variation.

Transgressive segregation: The appearance in F2 individuals with higher or lower intensity of

characters than the parents is called as transgressive segregation.

Modifiers/ Modifying genes: Group of genes, which enhance or reduce the phenotypic
expression of an oligogene. These genes have small, cumulative effect on the level of expression
of another gene.
Atavism, throw back or reversion: The reappearance of ancestral characters after several
generations because of recessiveness or other masking effect.

Quantitative characters or polygenic characters

Quantitative characters are traits which show continuous variation and influenced by
large number of genes in a cumulative fashion called as quantitative characters or polygeneic
character.
Whereas qualitative characters show discontinuous variation and governed by one or two major
genes or oligognes.

The inheritance of many different genes influencing the same character in a cumulative
fashion is called as multiple factor inheritance/ quantitative inheritance / polygenic inheritance.
The genetical studies of qualitative traits are called qualitative [Link] term polygene was
introduced by Mather in 1941. This term has wide usage in quantitative genetics.
Eg of Quantitative Inheritance :Many measurable traits viz., height, weight, yield and traits
such as intensity of colour and are governed by many genes (Fisher,1918).

Multiple factor hypothesis

Yule (1906) gave the theoretical explanation for the multiple factor hypothesis. According to him
“quantitative characters are controlled by many genes with cumulative effect without dominance
and would produce continuous variation”.
Causes of Genetic Variation: The Multiple-factor Hypothesis
We are used to thinking about the genetics of discrete plant phenotypes, for example tall
versus short peas in Mendel‟s experiments. In this example, variation (different alleles) at a
single gene contributes to a major phenotypic difference. However, for many traits of interest
(such as yield), it is often the case that a large number of genes, each of modest effect,
collectively contribute to the genetic variation. This is the multiple-factor hypothesis.
Early support for this multiple-factor hypothesis came from H. Nilsson-Ehle (1909), a
Swedish geneticist working on various cereal crops. Many of the characters that he examined
yielded 3:1 ratios in the F2 generation following the cross of two parental strains, consistent with
expectations for a single segregating locus with one allele completely dominant over the other.
However, there were some striking exceptions. For example, when red-seeded and white-seeded
wheat strains were crossed, the F1 progeny were identical in color (light red), but in some of the
F2 crosses, a ratio of 63 red:1 white seeds was observed. Nilsson-Ehle interpreted this to be the
result of the segregation of three independent factors, the initial parents being AABBCC and
aabbcc, all members of the F1 being AaBbCc and hence uniform in color, and the F2 consisting of
all possible genotypes, only one of which (aabbcc) gives rise to white seed. The probability of
obtaining an aabbcc offspring from an AaBbCc x AaBbCc cross is 1/64:
 From these results, Nilsson-Ehle arrived at two general conclusions. First, sexual
reproduction can produce a huge diversity of genotypes. Second, given this huge potential
diversity of genotypes, apparently new types appearing within a population may be the result of
rare segregants rather than new mutations.

1. Kernel Colour in Wheat

Nilsson-Ehle (1909) gave first experimental evidence for multiple
factor hypothesis in studies on the inheritance of kernel colour in wheat. He
crossed a strain of red kernel wheat plant with another strain of white kernel.
Grain from the F1 was uniformly red, but of a shade intermediate between the
red and white of the parental generation. This might suggest incomplete
dominance, but when F1 offspring were crossed among themselves, the F2
progenies showed five different phenotypic classes in a. ratio of 1 : 4 : 6 : 4:1.
Noting that 1/16 of the F2 was an extreme in colour as either of the parental plants (red or
white), they theorized that two pairs of genes controlling production of red pigment while
operating in this cross. Each gene was supposed to contain two alleles. One allele produces a
given quantity of the red pigment, while its counterpart did not produce any pigment. All alleles
were equally potent in the production or lack of production of pigment. If we symbolize the
genes for red with the capital letters R1 and R2 and their, alleles resulting in lack of pigment
production by r1 and r2: The F2 results of this cross as follows: or 1/16 Red: 4/16 Dark: 6/16
Medium: 4/16 Light: 1/16 White.

(R1 R2) (R1 r2) (r1 R2) (r1 r2)

(R1 R2) R1R1R2R2 R1R1R2 r2 R1r1R2R2 R1r1R2r2

DR MDR MDR MR
(R1 r2) R1R1R2r2 R1R1r2r2 R1r1R2r2 R1r1r2r2
MDR MR MR LR
 (r1 R2) R1r1R2R2 R1r1R2r2 r1r1R2R2 r1r1R2r2
MDR MR MR LR
(r1 r2) R1r1R2r2 R1r1r2r2 r1r1R2r2 r1r1r2r2
MR LR LR white

The F2 ratio in wheat -[Link]

This suggested that the seed colour in wheat is controlled by many genes which show
lack of dominance and have small and cumulative effects.
MAIN FEATURES
 One of the alleles of each colour gene produces seed colour and is called positive allele
denoted by capital letter (eg.) R1, R2 etc.
 The other allele of each colour gene does not produce any colour and is known as
negative allele denoted by corresponding small letter (eg.) r1, r2 etc.
 The heterozygote for colour gene is R1r1 is intermediate in colour between the two
homozygotes (R1 R1 and r1r1)
 Two positive alleles of a colour gene (R1 R1) produce twice the intensity of red colour of
that produced by a single positive allele (R1 r1)
 Each positive allele has a small, equal and additive ffect on seed colour.

Genotype and phenotype frequencies produced by two genes with cumulative effect on
seed colour in wheat

Genotypic No. of Phenotype

Genotype Frequency
ratio positive allele
R1R1R2R2 1 4 Dark Red 1

R1r1R2R2 2 3 Medium
Dark Red 4
R1R1R2r2 2 3
R1r1R2 r2 4 2 Medium
Red
R1R1r2r2 1 2 6
r1r1R2R2 1 2
R1r1r2r2 2 1 Light Red 4
 r1r1R2r2 2 1
r1r1r2r2 1 0 White 1

2. Skin Colour in Man

Another classical example of polygenic inheritance was given by Davenport (1913) in
Jamaica. He found that two pairs of genes, A-a and B-b cause the difference in skin
pigmentation between Negro and Caucasian people. These genes were found to affect the
character in additive fashion. Thus, a true Negro has four dominant genes, AABB, and a white
has four recessive genes aabb. The F1 offspring of mating of aabb with AABB, are all AaBb
and have an intermediate skin colour termed mulatto. A mating of two such mulattoes
produces a wide variety of skin colour in the offspring, ranging from skins as dark as the
original Negro parent to as white as the original white parent. The results of this cross are as
follows :
Parents: Negro Whit

AABB x aabb
Gametes: ↓
F1: Mulatto Mulatto
AaBb x AaBb

↓
F2 results:
Genotypic
Phenotypes Genotypes Frequency Phenotypic Ratio

Black (Negro) AABB 1 1

Dark AaBB 2 4

Dark AA Bb 2

Intermediate AaBb 4 6

Intermediate aaBB 1
Intermediate
AA bb 1

Light Aabb 2 4
Aa Bb 2
 Light
White aa bb 1 1

These results are clearly showing that A and B genes produce about the same amount of
darkening of the skin ; and therefore, the increase or decrease of A and B genes cause variable
phenotypes in F2 in the ratio of 1 Negro: 4 dark : 6 intermediate: 4 light : 1 white.

3. Height in Man
Skin colour in man is a rather simple example of polygenic
inheritance because only two pairs of genes are involved. The
inheritance of height in man is a more complex phenomenon
involving ten or more pairs of genes. The character of tallness is
recessive to shortness, thus, an individual having the genotype of more
dominant genes will have the phenotype of short stature. This trait is
highly influenced by a variety of environmental conditions. The height of adults ranged from 140
cm to 203 cm.
If one measured the height of a thousand adult men and the height of each is plotted against
height in centimeters and the points connected, a bell-shaped curve is produced which is
called curve of normal distribution and is characteristic of quantitative inheritance.

4. Other Examples
Likewise, if one measures the length of thousand sea shells of the same species, or counts
the number of kernels per ear in a thousand ears of corn, or weighs one thousand hen's egg, one
will find a normal curve of distribution in each case.

POLYGENES
The term polygene was introduced by Mather in 1941. This term has found wide usage
in quantitative genetics. A polygene or multiple factor or quantitative gene is a member of a
group of non-epistatic genes that interact additively to influence a quantitative trait. Advances in
statistical methodology and high throughput sequencing are, however, allowing researchers to
locate candidate genes for the trait. If such a gene is identified, it is referred to as a quantitative
trait locus (QTL). Whereas, oligogenes or major genes have larger effect on the characters and
shows discontinuous variation.

The phenotypic traits of the different organisms may be of two kinds viz., qualitative
and quantitative. The qualitative traits are the classical Mendelian traits of kinds such as (e.g.,
round or wrinkle seeds of pea); structure (e.g., horned or hornless condition in cattles); pigments
(e.g., black or white coat of guinea pigs); and so on. The organisms possessing qualitative traits
have distinct (separate) phenotypic classes and are said to exhibit discontinuous variations.
The quantitative traits, however, are economically important measurable phenotypic
traits of degree. The quantitative traits are also called as metric traits. They do not show clear
cut differences between individuals and forms a spectrum of phenotypes to cause continuous
variations. The quantitative traits may be modified variously by the environmental conditions
and are usually governed by many genes (perhaps 10 or I00 or more), each contributing such a
small amount of phenotype that their individual effects cannot be detected by Mendelian
methods but by only statistical methods.

S. No. Qualitative characters Quantitative characters

1. Controlled by oligogenes (one or Controlled by polygenes (each with small
two genes) or major genes cumulative effect) or minor genes
2. Discontinuous variation Continuous variation
3. Less influenced by environment Much influenced by environment
4. Concerned with individuals mating Concerned with population of organisms
and their progeny consisting of all possible kinds of mating.
5. Exhibit high heritability Exhibit low heritability.
6. Analysis by making counts and Analysis by mean, variance and covariance
ratios
7. Classified into clear cut groups Cannot be classified into clear cut groups

Characteristics of Quantitative Inheritance

i. Each polygenic character is controlled by several genes and has cumulative effect.
ii. Polygenic traits are highly sensitive to environmental changes. Polygenic characters
exhibit continuous variation. Hence, they cannot be classified into clear cut groups.
iii. Effect of individual gene is not easily detectable in case of polygenic characters and,
therefore, such traits are also known as minor gene characters.
 iv. The statistical analysis of polygenic variation is based on means, variances and co-
variances, whereas the discontinuous variation is analysed with the help of frequencies
and ratios.
v. Thus, polygenic characters are studied in Quantitative genetics and oligogenic
characters in Mendelian genetics.
vi. Generally the expression of polygenic characters is governed by additive gene action,
but now cases are known where polygenic characters are governed by dominance and
epistatic gene action.
vii. In case of polygenic characters, bio-metric measurements like weight, duration,
strength, etc. are possible
viii. Transgressive segregants are possible from the crosses between two parents for a
polygenic character.
ix. The transmission of polygenic characters is generally low because of high amount of
environmental variation. On the other hand, oligogenic characters exhibit high
transmission because there is little difference between the genotype and phenotype of
such character.
x. Polygenic inheritance : Additive effects of two or more genes on a character

Transgressive segregation
 In genetics, transgressive segregation is the formation of extreme phenotypes,
or transgressive phenotypes, in segregating populations compared to phenotypes observed in
the parental lines. The appearance of these transgressive (extreme) phenotypes can be either
positive or negative in terms of fitness. The appearance in F2 individuals falling outside the
parental range in respect to some character is called transgressive segregation.
Eg Parents AA BB cc dd ee x aa bb CC DD EE

F1 Aa Bb Cc Dd Ee
F2 aa bb cc dd ee and AA BB CC DD EE

 Transgressive segregation results due to fixation of dominant and recessive genes in

separate individuals.
 Such segregation occurs when the parents are intermediate to the extreme values of the
segregating populations.
 The superior plants are produced by an accumulation of favourable genes from both the
parents as a consequence of recombination.
Modifiers/ Modifying genes
 Group of genes, which enhance or reduce the phenotypic expression of an oligogene.
 These genes have small, cumulative effect on the level of expression of another gene.
 They do not effect the concerned character on their own, they seem to modify the
expression of the concerned major gene.
 Expression of many oligo gene is affected by modifying genes.
 Modifying genes produce a continuous variation in a character governed by an
oligogenes.
Eg: Self Incompatibility and male sterility genes in plants, genes for disease resistance
Instead of masking the effects of another gene, a gene can modify the expression of a
second gene. In mice, coat color is controlled by the B gene. The B allele conditions black coat
color and is dominant to the b allele that produces a brown coat.
 The intensity of the color, either black or brown is controlled by another gene, the D
gene.
 The dominant D allele controls full color whereas the recessive d allele conditions a
dilute or faded expression of the color expression at the B gene.
In a cross is made among mice that are BdDd, the following phenotypic
distribution will be seen:
9 B_D_ (black)
3 B_dd (dilute black)
3 bbD_ (brown)
1 bbdd (dilute brown)
The D gene does not mask the effect of the B gene, rather it modifies its
expression.
Lecture 22-Linkage - Coupling and repulsion; Experiment on Bateson and Punnet

Linked genes : Two or more genes present on the same chromosome and they do not exhibit
independent assortment, they are said to be linked
Linkage: The phenomenon of transmission of linked genes is called linkage.
Coupling : The tendency of both dominant alleles or both recessive alleles to pass together to the
same gamete.
Repulsion : The tendency of both dominant or both recessive alleles to repel each other, so that
gametes with one dominant and one recessive will form more frequently.

LINKAGE
Every individual organism bears several heritable characters which are represented by the
innumerable genes present on the chromosomes. During meiosis, the chromosomes move into
the gametes as units, all the genes present on any given chromosome will segregate as a group
and move together from generation to generation. This tendency of the genes located on the
same chromosome, to stay together in hereditary transmission, is known as linkage. The
genes located on the same chromosome are called as linked genes.
No Linkage Linkage
Independent Assortment Without Recombination

Prediction of linkage
• Linked genes do not exhibit independent assortment
• Large deviations from 1 : 1 : 1 : 1 in the testcross progeny of dihybrid experiment is used
as a first evidence for linkage
 • Number of recombinant progeny is significantly less than expected from Mendel‟s
Second Law

Symbol of linked genes

While representing linked gene, the two homologous chromosomes are indicated by two
horizontal links.

Coupling and repulsion

Coupling is a condition of linked inheritance in which an individual heterozygous for two
pairs of genes receives the two dominant alleles from one parent and the two recessive alleles
from the other parent.
Repulsion is the condition in linked inheritance, in which an individual heterozygous for
two pairs of linked genes receives the dominant allele of one pair gene and the recessive allele of
the other pair of gene from one parent and the reverse from the other parent

The principle of linkage was discovered by Bateson and Punnet in 1906 in the sweat
pea plant, Lathyrus odoratus. However, linkage, as a concept was put forth by Thomas Hunt
Morgan in 1910 based on his experiment on Drosophila melanogaster.
Bateson and Punnett (1905 -1908) formulated “hypothesis of coupling and repulsion” to
explain the unexpected F2 results of a dihybrid cross.
  The experiment indicates that two genes are located on the same chromosome and said to
be linked.
 The linked genes tend to pass on to the same gamete and responsible for the appearance
of more parental combinations in the progeny.
 Small number of recombination due to some amount of recombination between linked
genes.
 The [Link] test cross ratio clearly indicated that there was a tendency in the dominant
alleles (BL-Purple long) to pass together to the same gamete.
 Similar was the case with recessive alleles (bl-red round).

The [Link] test cross ratio clearly indicated that there was a tendency in the dominant alleles
(BL) and both recessive allele (bl) to pass together to the same gamete. This tendency of
dominant and recessive alleles to inherit together was explained as “gametic coupling” by
Bateson and Punnett.
 This experiment explains the repulsion phase. The tendency of both dominant and both
recessive alleles to repel each other, so that gametes with Bl and bL are formed more
frequently.
Bateson and Punnet’s Conclusion
 The alleles, which come from the same parents, tend to enter the same gametes.
 The alleles which come from different parents tend to enter different gametes.
 When the genes are linked, greater than 50% of progeny with parental phenotypes and
less than 50% of progeny with recombinant phenotypes will occur.

Linkage in maize
'C' for coloured aleurone is dominant over 'C' colourless Sh for Full endosperm is
dominant over 'sh' shrunken.
F2 did not show 9: 3: 3: 1 ratio. There were greater number of colour full, colour
shrunken (parental types) than colourfull shrunkern , colour full, If two character considered
separately, they segregate 3 : 1
 i.e . Colour - 7500 Full - 7500
Colourless - 2500 Shrunken - 2500
The large deviation of the observed F2 population form the excepted segregation is
therefore not because the members of each pair of alleles do not segregate from each other but
because of the separation in one pair of alleles is not independent of the separation in the other
pair of alleles.

Segregation of two pairs of genes on two pairs of chromosomes

Let us suppose that, gene 'C' is located on chromosome number 9 and 'S' on chromosome
number 10 of maize. The segregation of chromosome bearing C and c is entirely independent of
segregation of chromosome bearing S and s. So four type of gametes Cs, Cs, eS, eS are formed
in F1 and F2 normal dihybrid ratio [Link] and test cross [Link]
Segregation of two pairs of genes on one pair of chromosomes
Let us suppose that, two genes C and S are located on chromosome No. 9 during meiosis
only 2 gametes will be formed CS and cs gametes. So, Genes C and S situated on same
chromosomes are said to be linked. Linkage is the association of character in inheritance due to
fact that genes determining them are physically located on the same chromosomes.
Lec 23 -Chromosomal theory of linkage of Morgan – Complete and incomplete linkage,
Linkage group

Detection of Linkage
Compare the number of individuals observed in each class with those expected on the
basis of independent assortment and then to test the deviation between these two values by chi-
square test.

Linkage and Recombination

Exchange of corresponding segments between the non-sister chromatids of homologous
chromosomes leading to recombination of linked genes and was first observed by Belgian
cytologist Janssens in 1909.
[Link] has related linkage to the segregation of homologous chromosomes and
occurrence of crossing over during first meiotic division. Morgan‟s interpretation of linkage is
known as “The chromosomal theory of linkage”. Results of crosses involving linked genes in
fruit fly Drosophila melanogaster- Published in 1911. He has studied two characters involving
two different genes.
 One gene affects the body colour [grey body (b+) which is dominant over black body
(b)].
 The second gene affects the phenotype of the wing [long wing (s+) which is dominant
over vestigial wing or short wing (s)].

Cross I long wing x short wing

grey bodied flies black bodied flies
s+b+/s+b+ sb/sb
F1 long wings and grey bodies (s+b+/sb) x sb/sb
Test cross 82 per cent (41 each)- parental combination of traits
18 per cent (9 each)- recombinant combinations
 Cross II
Long wing x short wings
black bodies grey bodies
s+b/s+b sb+/sb+
F1 long wings and grey bodies (s+b/sb+) x sb/sb
Test cross 82 per cent - parental phenotypes
18 per cent - recombinant phenotypes

 The genotype of a heterozygote (s+sb+b) of this type is frequently written as s+b+ / sb.
This arrangement of wild type (dominant) form of two genes in a heterozygote is called
the coupling phase or cis – configuration (AB/ab)
 The alternative arrangement, illustrated in cross II where each homologue contains one
mutant gene and one wild type gene (s+b / sb+) is called the repulsion phase or trans
configuration. (Ab/aB)
 Test cross progeny of the F1 female flies contain very different frequencies of the four
phenotypic classes in the two cases.
 41 per cent of the progenies in cross I are wild type (have long wing and grey bodies); in
cross II only 9 per cent are wild type.
 Allelic forms of the two genes that are present together on the homologous chromosomes
of the parent tend to remain together on the chromosome of the progeny.
 In cross I, the F1 flies carried the wild type forms (s+ b+) of the two genes on one
homologue and the mutant forms (sb) on the other homologue.
Main features of Chromosome theory of linkage

 Genes located on the same chromosome are inherited together. They are said to be linked.
 Genes are present in the chromosomes in a linear order.
 The distance between the linked genes determines the degree of linkage. Closely located
genes show strong linkage and genes widely part show weak linkage.
 Only those genes located in different chromosome show independent assortment.
 Genes that are located far apart on the same chromosome would also assort
independently.
Linkage studies revealed the following
 Genes that assort at random are non linked genes. Genes that do not segregate at random are
linked genes.
 Linked genes are arranged in a lines fashion on the chromosome. Each linked gene has a
definite and constant order in its arrangement.
 The distance between the linked genes determines the degree of strength of linkage. Closely
located genes show stronger linkage that the widely located genes.
 Linked genes do not always stay together, but are often exchanged reciprocally by cross over.
Types of linkage
1. Based on crossing over
Complete Linkage
The genes closely located in the chromosome show complete linkage as they have no
chance of separating by crossing over and are always transmitted together to the same
gamete and the same offspring. Thus, the parental combination of traits is inherited as
such by the young one.
Incomplete Linkage
The genes distantly located in the chromosome show incomplete linkage because they
have a chance of separation by crossing over and of going into different gametes and
offspring.
2. Based on genes involved
Coupling linkage – AB/ab
Repulsion linkage –Ab/aB
3. Based on chromosome involved
1. Autosomal linkage – somatic chromosomes
2. X-chromosomal linkage – Sex chromosomes

Linkage Group
The number of linkage groups will be equal to the haploid number of chromosomes
which the species possess. Thus maize with 10 pairs of chromosomes has 10 linkage groups.
Significance of linkage
1. Effect in selection- Linkage between two or more loci controlling different desirable
characters is advantageous in selection programme.
2. Effect in genetic variance : Linkage in repulsion phase in segregating population causes
upward bias in estimate of dominance variance and downward bias in the estimate
additive genetic variance.
3. Effect in genetic correlation : Linked traits show high genetic correlation and co-
heritability.
4. Effect on genetic improvement: Linkage between genes controlling two desirable traits
help in simultaneous improvement of both characters.

Linked Genes on the Same Chromosome Exhibit Distorted Mendelian Ratios

It was not long from the time that Mendel's work was rediscovered that new anomalous
ratio began appearing. One such experiment was performed by Bateson and Punnet with sweet
peas. They performed a typical dihybrid cross between one pure line with purple flowers and
long pollen grains and a second pure line with red flowers and round pollen grains. Because they
knew that purple flowers and long pollen grains were both dominant, they expected a typical
[Link] ratio when the F1 plants were crossed. The table below shows the ratios that they
observed. Specifically, the two parental classes, purple, long and red, round, were over
represented in the progeny.
Phenotype Observed Expected
Purple, Long (P_L_) 284 215
Purple, round (P_ll) 21 71
Red, Long (ppL_) 21 71
Red, round (ppll) 55 24
Total 381 381

At the time of these experiments, Bateson and Punnett were not able to develop an
acceptable hypothesis. The best explanation they posed was that in some manner the phenotypic
classes (alleles) in the parents were coupled, and they did not sort independently into gametes
 as predicted by Mendel's second law.

Proof those genes on the same chromosome can at times be inherited as blocks was given
by T.H. Morgan with Drosophila. Morgan crossed red eye, normal wing flies (pr+pr+ vg+vg+)
with purple eye, vestigal wing (prpr vgvg) flies. During meiosis, four different F1 gametes are
produced. The parental gametes are developed without any processing. The recombinant
gametes though occur by a process called crossing over. (The X between the two F1
chromosomes represents the crossing over event.).
Morgan performed a testcross by crossing prpr vgvg flies to F1. The testcross is powerful
because it allows you to follow the meiotic events in one parent because all of the gametes from
the test cross parent are homozygous recessive. For this example, the testcross genotype is pr vg.
Therefore the testcross progeny will represent the distribution of the gametes in the F1.
Remember that a testcross to F1 derived from a dihybrid cross gave a [Link] ratio. But this is not
what Morgan observed. The following table shows the result of this test cross.

F1 Gamete Testcross Distribution Gamete Type

pr+ vg+ 1339 Parental
pr+ vg 151 Recombinant
pr vg+ 154 Recombinant
pr vg 1195 Parental
 These results confirm the Bateson and Punnett hypothesis that two genes do not always
assort independently. A further confirmation experiment was performed by Morgan when he
crossed red eye, vestigal wing flies and purple eye, normal wing flies. Whereas in the first cross,
the two dominant alleles and two recessive alleles were on the same chromosome the F1, in the
is cross a dominant allele was on the same chromosome as a recessive allele. The term for the
first chromosomal arrangement of the F1 is called coupling, whereas the second arrangement is
called repulsion. Another set of terms to describe these arrangements are cis and trans,
respectively. The following shows the chromosomal arrangement for the cross of two parents in
repulsion.

As with the first cross, Morgan test crossed these F1 flies. The following table shows the
distribution of these F1 gametes.

F1 Gamete Testcross Distribution Gamete Type

pr+ vg+ 157 Recombinant
pr+ vg 965 Parental
pr vg+ 1067 Parental
pr vg 146 Recombinant

It was expected that both the coupling and repulsion crosses would yield [Link] ratios.
How can we determine if the results deviate from this ratio. As with any ratio, we can use the
chi-square test to determine if the observed results fit or deviate from the expected ratio. The
two tables below show the results for the chi-square for the two crosses.
Coupling Cross Chi-Square Test

F1 Gamete Observed Expected (O-E)2/E

pr+ vg+ 1339 709.75 557.9
pr+ vg 151 709.75 439.9
pr vg+ 154 709.75 435.2
pr vg 1195 709.75 331.8
Total 2839 2839 X2=1764.8

Repulsion Cross Chi-Square Test

F1 Gamete Observed Expected (O-E)2/E
pr+ vg+ 157 583.75 312.0
pr+ vg 965 583.75 249.0
pr vg+ 1067 583.75 483.3
pr vg 146 583.75 328.3
Total 2335 2335 X2=1372.6

It is quite clear that both of these large chi-square values indicate that neither of these ratios fit
the [Link]
 Lecture 24-Crossing over – significance of crossing over; cytological proof for crossing
over - Stern’s experiment; Factors controlling crossing over.
Crossing over: It is the mutual exchange of the chromosome segments between non-sister
chromatids of a homologous pair of chromosomes during pachytene stage of meiosis I,
producing new combinations of genes.
Cross over chromatids: The chromatids resulting from the interchange of segments
Non-crossover parental chromatids :The chromatids which does not involved in crossing over
and remain intact.
Crossing over between non-sister chromatids of a homologous pair of chromosomes
produces new combinations of genes.

Main features of crossing over

i. Takes place at four strand stage (tetrad) during pachytene stage
ii. Occurs between non-sister chromatids of homologous chromosomes
iii. Produces new combination of genes between linked genes
iv. Value of crossing over or recombination may vary 0-50%
v. The number of crossing over depends upon the length of the chromosome.
vi. The longer the length, the higher the percentage of crossing over.
vii. When the genes are closely located the chance for crossing over is lesser
viii. The number of crossing over between two genes is represented by percentage of crossing
over
 ix. The percentage of crossing over or recombination is directly proportional to the distance
between two genes. The percentage of crossing over is the expression of the number of
recombinants to the total number of offspring in percentage

Total no of recombinants
Percentage of crossing over= ________________ x 100
Total [Link] offspring in F2
x. The percentage of crossing over is also called frequency of crossing over
Thus percentage of crossing over = Frequency of crossing over
= Percentage of recombination
= Frequency of recombination

Types of crossing over

1. Single crossing over (Two strands) : When there is a single crossing over in a
homologous chromosome, it is called single crossing over
2. Double crossing over (2,3, or 4 strands): When there are two cross over between non-
sister chromatids of homologous chromosome.
3. Multiple crossing over : When there are many cross over in a homologous chromosome,
it is called multiple crossing over. It occurs in very low frequency. If the genes are on the
same chromosome, multiple crossovers can occur.
Significance of Crossing Over
1. Linear arrangement of genes: Crossing over clearly illustrates the linear arrangement of
genes on the chromosomes.
2. Chromososme Maps: The frequency of crossing over is very useful to construct the
chromosome maps.
3. Recombination: Crossing over produces new combination of genes.
4. Variations: Crossing over leads to genetic variation which is the raw material for
evolution.

Cytological Proof of crossing over

Cytological markers: Cytologically distinguishable features which can differentiate the two
members of a homologous pair of chromosomes.
1. Stern’s Drosophila Experiments
In 1931, a few weeks after publication of similar experiments by Creighton and
McClintock in Zea mays, Stern reported work with Drosophila.
Two linked characters – Colour and shape of the eye
 Colour : Red eye (+) –wild type is dominant and carnation eye is recessive (car).
 Shape : Bar eye is dominant (B) over wild type- round eye (+) which is recessive.
Male had normal X chromosome which carries allele for carnation and round eye car +
and an Y chromosome. The female had 2 cytologically distinct X chromosomes. One X had a
portion of the Y chromosome attached to it, making it longer than normal. This one had wild
type allele (+ +) for both genes. The other X was shorter than normal, and had the car and B
alleles on it.
This female having red bar eyes is crossed with a double recessive male
having carnation round eyes (car,+).
  In the absence of crossing over, only two types of female gametes are produced. One
type having broken X containing car and B genes; the other type having the X with a
piece of Y chromosome attached containing + and +.
 Crossing over between two X chromosome produces two more types of gametes.
 One type having car and + in a normal size X and the other type having + and B on a
broken X with a piece of Y chromosome.

The observed results require the above types of gametes to be produced by [Link] four
types of gametes, after fertilization will produce four types of offspring and they are
[Link] bar 2. Red non-bar 3. Carnation round and 4. Red bar
The X chromosomes of these four types are identified. Each offspring has the expected
chromosomal configuration. This experiment proves that exchange of chromosomal material
takes place between the homologous chromosomes.

F1 progenies
 The key observation was that in every case of genetic recombination, there had also
been recombination of the cytological features. Genetic recombination was always
accompanied by recombination of the cytological features. The same type of results had been
found in the corn experiments.

2. Corn Experiments
Creighton and McClintock (1931) worked with corn (Zea mays) plants in which the two
chromosomes under study differed cytologically. The study used a corn strain heterozygous
for two genes on chromosome 9. One gene determines seed color (C for colored seeds, c for
colorless). The other gene is involved in starch synthesis. The wild-type allele (Wx) produces
amylose, and the combination of amylose and amylopectin forms normal starch in a corn seed.
The waxy mutant (wx) lacks amylose, and has waxy starch containing only amylopectin.
In this corn strain, the appearance of each chromosome 9 homolog correlated with its
genotype: One chromosome 9 had the genotype c Wx, and a normal appearance. Its homolog
had the genotype C wx, and cytological markers at each end of the chromosome. The end
near the C locus had a darkly staining knob, and the other end, nearer the wx locus, had a
translocated piece of chromosome 8.

When testcrossed, recombinant phenotypes were evident, and could be correlated with
cytological features. Whenever the genes had recombined, the cytological features had also
recombined. In the parental (non-recombinant.) type progeny, no exchange of cytological
markers was evident. This was direct evidence of physical exchange between homologs
resulting in genetic recombination
These two studies convinced the field of genetics that physical exchange of chromosome
segments was the basis for recombination of linked genes.
Factors Affecting Crossing Over
1. High temperature increases the frequency of crossing over.
2. X-ray increases the frequency of crossing over.
3. The frequency of crossing over decreases with increasing age in female Drosophila.
4. Some genic mutation decreases the frequency of crossing over.
5. Crossing over is less frequent near centromere and the terminal region of the chromosomes.
6. Inversions and translocations of chromosome segments suppress the crossing over.
7. Crossing over at one point of chromosome reduces the frequency of another crossing over in
its adjacent region. This phenomenon is known as interference.
7. Sex of the organism: Heterogametic sex shows relatively lower recombination frequencies
8. Distance between genes: Greater the distance, higher the chance of crossing over.
9. Nutrition: Recombination was affected by the presence of Calcium and Magnesium in the
diet.
10. Cytoplasmic genes: In pearl millet, Tifton MS cytoplasm reduce recombination
Lec 25 : Strength of linkage and recombination; Two point and three point test cross
Recombination frequency: The ratio of number of recombinants to the total number of
progeny
Two point test cross: It is a test cross which involves a dihybrid and a double recessive parent
to determine the crossover percentage between two linked genes.
Three point test cross: It is a test cross which involves a trihybrid and a double recessive
parent to determine the distance between the three genes and the order in which they are located
on the chromosome.

Recombination frequency
Recombination frequency/ Crossing over frequency (%) =
No. of recombinants
--------------------------- ×100
Total progeny
Used in assessing the relative distance between genes on a genetic map
Chiasma frequency
A pair of synapsed chromosome is known as tetrad. Every tetrad has atleast one
chiasma. In the long chromosome, number of chiasmata will be more. The farther apart two
genes are located on the chromosome, the greater the opportunity of chiasma formation between
them. Frequency with which chiasma occurs has a characteristic probability. It is useful in
predicting proportion of parental and recombinant gametes. The percentage of crossover
(recombinant) gametes formed by a given genotype is a direct reflection of chiasma frequency.
When a chiasma formed between two gene loci, only half of the meiotic products will be
crossover type.
Therefore, Chiasma frequency (%) = Twice the frequency of crossover frequency
=2(crossover frequency) or
Crossover frequency (%) = ½ (Chiasma frequency )
Strength of linkage and recombination
• Strength of linkage depends on distance between two genes.
• Closely linked genes – strong linkage
• Widely located genes-weak linkage
• Confirmed by test cross (Heterozygous F1 with recessive parent)
• Parental types- Complete linkage; recombinant types- Incomplete linkage
• More % of Parental types – strength of linkage is high
• More % of recombinant types – strength of linkage is low
Limits of recombination
• If two loci are far apart on the same chromosome, probability of chiasma formation is
100 per cent. Then 50 % of the gametes will be recombinant and 50% will be parental
type.
• Recombination between linked genes not > 50% even when multiple crossing over
occurs

Two-Point Testcross
It is done to determine the recombinant frequency between two linked genes. The percentage of
crossing over between two linked genes is calculated by test crosses in which a F1 dihybrid is
crossed with a double recessive parent. Crossing over at two points is two point test cross
involving two pairs of genes.
For eg., a dihybrid having the genotype AC/ac is test crossed with double recessive parent
(ac/ac). In F2 test cross progenies
37% dominant genes at both gene loci (AC/ac) (PT),
37% recessive genes at both gene loci (ac/ac) (PT),
13% dominant genes at 1st gene loci and recessive gene at second gene loci (Ac/ac),
13% recessive genes at 1st gene loci and dominant gene at second gene loci (aC/ac)c),

Construction of Chromosome map in Drosophila Using Two-Point Testcross

In Drosophila, the chromosome map is constructed with the help of test cross. In Drosophila,
grey colour is dominant over black colour; and the long wing is dominant over vestigial wing.

The F1 female hybrid is test crossed, four types of individuals are formed. Out of four
types, two types are parental type (b+v+ /bv & bv/bv) and other two are non parental type
(b+v/bv & bv+/bv) due to crossing over. Non – parental type is 17%. So the percentage of
crossing over is equivalents to 17%. The distance between the two genes (b+:v+) is equivalent to
the percentage of crossing over or percentage of non parental combination. So the distance
between the gene b+:v+ is equivalent to 17 cM or mapunit.

Percentage of non parental combination = 17%

So the percentage of crossing over = 17

So the distance between the Gene b+:v+ = 17 cM

 In another experiment, the F1 female grey red is test crossed with black cinnabar. The
experiment shows 9% non parental combination individuals. So the distance between the Gene
b+ & cn+ is equivalent to 9 map unit or 9 cM.
In the same way the F1 female red long is test crossed with cinnabar vestigial. The
experiment shows 9.5% non-parental combination individuals. So the distance between the gene
v+ & cn+ is equivalent to 9.5 map unit.

According to the first experiment the distance between b+ & v+is equivalent to 17 map
unit. But the second and third experiment show 18.5 map units between the two genes. To find
out the actual reason for this difference in the distance, a three point cross has to be
conducted.

Three Point cross

Three point test cross or trihybrid test cross involving three genes gives relative
distance between these genes, and also shows the linear order of these genes on the
chromosome. Such test cross may be carried out if three points or gene loci on chromosome
can be identified by marker genes. In addition to the genes A and C, a third marker gene B is
located in fairly closed proximity in the same linkage group, all three markers may be used
together in conducting a more precise analysis of the map distance and relative position of three
points
In the three point cross, all the three pairs of genes are considered in the experiment.
When the F1 hybrid female is test crossed, they produce 8 different types of individuals. Out of
8 types, two types are parental and the remaining six are non-parental.
Female Male

Parent : Black cinnabar Vestigial Grey Red Long

b cn v / b cn v X b+ cn+ v+/Y

Normal
F1 :
b+ cn+ v+/ b cn v

Test cross :
Female Male
Normal Recessive
b+ cn+ v+ / b cn v x b cn v / Y
Test cross progenies
Total progenies: 800
1. Parental combinations due to linkage = 658 = 82.25%
2. Non parental combination:
Single cross over:

a) Between b+ and cn+

1. Black red long b cn+ v+/b cn v = 36 = 70

2. Grey cinnabar vestigial b+ cn v/b cn v = 34

b) Between cn+ and v+
1. Grey red vestigial b+ cn+ v / b cn v = 35
=66
+
2. Black cinnabar long b cn v / b cn v = 31

Double cross over

1. Grey cinnabar long b+ cn v+ / b cn v =4 =6

2. Black red vestigial b cn+ v / b cn v =2

7
Total c.o between b+and cn+ =70 +06 = 76 = 9.5%
(includes double c.o)
6
Total c.o between cn+ and v+ =66 +66 = 72 = 9.0%
(includes double c.o)
7
Total c.o. between b+ and v+ =76 +672 = 148 = 18.5%
(includes double c.o)
7
C.o between b+ and v+ =70 +066 = 136 = 17%

(double c.o not included)

From these results, it is concluded that the gene cinnabar (cn+) lies about half – way
between the genes for black body colour and vestigial wings (b+ and v+). The total amount of
crossing over between black body and vestigial wing is 18.5% rather than the 17% expected on
the basis of the first cross. The discrepancy (18.5 – 17 = 1.5) just noted, arises because of the
occurrence of double crossing over, i.e. two cross overs occurring simultaneously between these
two loci.
As a final check on these results, it would be well to make a trihybrid or three point
cross using all three pairs of genes at once. When pure recessive flies are crossed with normal
flies, all the F1 flies are normal phenotypically. When the F1 females are back crossed to triple
recessive males, eight phenotypes are obtained.
From the data obtained the relative position of the genes can be calculated. The distance
between b+ and cn+ is 9 units, the distance between cn+ and v+ is 9.5 units; the v+ gene could
be to the right of cn+ locus or to the left. If the first order (cn+ v+) is correct, then the distance
between b+ and v+ is 17 units. This small discrepancy is due to double crossing over. Based
upon the above data, the three genes can be mapped as follows:
--------------------------------------------------------
b+ 9cM cn+ 9.5cM v+
Lec 26: Double cross over, interference and coincidence; genetic map, physical map
Double cross over: Formation of two chiasmata between the two loci.
Interference: The phenomenon of occurrence of a crossing over at one point reduces the
chance of occurrence of another crossover in its adjacent region.
Coincidence: The ratio of observed double crossover to the expected double crossover.
Genetic map: Line diagram which depict the position of various genes on the chromosomes
and recombination frequency between them. The map distance between the two genes is based
on cross over percentage. It is also known as linkage map or chromosome map.
Physical map: It shows the physical distance between the genes on the chromosome based on
base pairs. Physical maps are used to help scientists identify and isolate genes.

Types of double crossover

[Link]-strand double crossover: Result of two chiasma between the two loci involving the
same two non-sister chromatids. These two events cancel each other out and result in parental or
non-recombinant progeny.
[Link]-strand double crossovers: Result when one chromatid crosses over with two other
chromatids. The result is two parental and two recombinant progeny, the same as a single
crossover event.
Four-strand double crossover : is a single crossover between two chromatids and then a
second single crossover between the remaining chromatids. The result is that all four products
are recombinant.
Interference
Normally the double crossing over frequency is very low. Because the crossing over and
chiasma formation in the homologous non sister chromatids interferes with the crossing over
and chiasma formation at other points nearby. The phenomenon of occurrence of a crossing
over at one point reduces the chance of occurrence of another crossover in its adjacent region.
This was discovered by Muller (1911). The interference is inversely proportional to the
crossing over percentage. The interference is maximum over a short distance and decreases as
the distance increases.

Coincidence
The coincidence is an inverse measure of interference. It is measured as a ratio of
observed number of double cross over to the expected number of double cross over.

Observed number of double cross over

Coincidence = Expected number of double cross over

Interference = 1 - coefficient of coincidence. The values are inversely related.

If the actual number of double cross over is zero, then coincidence is zero and
interference is complete. If the actual number of double cross over is the same as the expected
number, coincidence is said to be one, and interference is nil. It ranges from 0 to 1.
Types of map
1. Physical map
2. Chromosome map/genetic map/linkage map/crossover map
1. Physical map: A physical map of a chromosome or a genome which shows
the physical locations of genes and other DNA sequences of interest. Distance is measured
based on the actual number of nucleotide pairs between loci. For the human genome, the
lowest-resolution physical map is the banding patterns on the 24 different chromosomes; the
highest-resolution map is the complete nucleotide sequence of the chromosomes.

2. Linkage map / Genetic map / Cross over map / Chromosome Map

The linkage maps are also referred as cross over maps since they are sketched by the amount of
crossing over. The percentage of crossing over is directly proportional to the distance of the
alleles showing crossing over in the chromosome.
The chromosomes maps are the graphic representation of the genes in a chromosome. The
percentage of crossing over is calculated using test crosses. In mapping the genes, a unit of
distance is used and it is called as map unit or Morgan unit (cM). Genetic and physical maps
illustrate the arrangement of genes and DNA markers on a chromosome.
Chromosome mapping or Genetic Mapping: The process of assigning genes on the
chromosome is known as chromosome mapping. It is a method used for determining the
location of and relative distances between genes on a chromosome.
Two aspects of genetic mapping
i. The determination of linear order
ii. The determination of relative distance between the genetic units
The first chromosome map was made in 1913 by [Link] and Sturtevant and soon after
additional maps was made by Bridges and others. Drosophila is the earliest material used by the
scientists, for constructing the maps.
Concept of Genetic Map (Linkage map)
In an individual heterozygous at two loci, there are two arrangements of alleles:
1. Cis (coupling) arrangement has both wild type alleles on one homologous chromosome,
and both mutants on the other (e.g., w+ m+ and w m).(AB /ab)
2. Trans (repulsion) arrangement has one mutant and one wild-type on each chromosome
(e.g., w+ m and w m+) (Ab/ aB)
3. A crossover between homologs in cis arrangement results in a homologous pair with the
trans arrangement (Ab/aB). A crossover between homologs in the trans arrangement
results in cis homologs (AB /ab).
4. Cross over frequency for linked genes (measured by recombinants) is characteristics for
each gene pair. The frequency stays the same, whether the genes are in coupling or in
repulsion.
5. An 1% crossover rate is a genetic distance of 1 map unit (mu). A map unit is also called
as centiMorgan (cM).

Procedure for the chromosome/genetic mapping

1. Determination of linkage groups
Before starting the genetic mapping of the chromosomes of a species, one has to know
the exact number of chromosomes of that species. Linkage group is a set of genes at
different loci on the same chromosome that except for crossing-over tend to inherit as a
unit in meiosis instead of undergoing independent assortment. All the genes which are
located on a single chromosome form one linkage group. The total number of linkage
groups in an organism corresponds to the haploid number of chromosomes, i.e., to the
 number of chromosome pairs. In Drosophila, 2n = 8; n = 4. This means that the haploid
number of chromosomes in Drosophila is 4. Therefore, there are 4 linkage
groups in Drosophila.

2. Determination of map distance

After knowing total number of genes in each linkage group of a species, the relative
distances between each linked gene have to be determined. The distance between two given
genes is calculated using two point test cross and three point test cross based on the % of
crossing over.
Eg. If % of crossing over is 1%, the map distance between two linked genes is one unit. If
mean number of chiasmata is known for a chromosome pair the total length of map for that
linkage group may be predicted.

3. Determination of gene order

After determining the relative distance between the genes of a linkage group, it becomes
easy to place genes in their proper linear order.
For eg. When the % of crossing over between two genes is 5, then the distance is 5 units.
For example five genes A, B, C, D and E are to be plotted on a chromosome. If cross over
results indicate that genes A and E have the highest percentage of crossing over, it means that
these should be placed at the maximum distance. In this example, the gene A can be taken as a
starting point in the chromosome and can be represented by O.
Now if the gene A and B exhibit 7% crossing over, the gene B can be placed on the
chromosome at a distance of 7 units. If the gene C shows 8% crossing over with gene B and
about 15% crossing over with gene A, it can be plotted on the chromosome at a distance of 15
units from gene A. Similarly if gene A and E exhibit 20% and 30% crossing over with gene D
and 5% and 10% with gene C these, are located on the chromosome 5 and 10 units away from
the gene C respectively.
Three point test cross is used to determine the distance between the genes and the order of genes
on the chromosome.
Eg. ABC/abc are three linked genes located on two different chromosomes in F1 of a cross
between AABBCC and aabbcc parents.
Three point test cross produced eight different phenotypic classes. Eight classes are
identified by phenotypic frequencies and alteration of gene sequence in the genotype. Parental
types have the maximum phenotypic frequencies and double cross over types produce least
frequencies.
Summary of the results of a three point test cross between ABC/abc x abc/abc abc

Genotypic classes Phenotypic classes Assumed frequencies Remarks

ABC/abc ABC 349

Parental types
abc/abc abc 360
Abc/abc Abc 114 Single crossover between
A and B
aBC/abc aBC 116
ABc/abc ABc 128 Single crossover between B
and C
abC/abc abC 124
AbC/abc AbC 5 Double crossover between
A and C
aBc/abc aBc 4
Total 1200

Single crossover between A and B will alter the position of two genes viz., B and C. Single
crossover between B and C will alter the position of only one gene viz., C and double crossover
between A and C will alter the position of only middle gene, i.e. B
The recombination percentage or unit distance between genes is worked out by
calculating the crossing over percentage between different genes.
Suppose number of crossover progeny between genes A and B is P
Single cross over between genes B and C is Q
Double cross over between genes A and C is R and total progeny is T,
Then, recombination (%)

1. Between genes A and B = P+R 114+116+9

------ x 100 = --------------x100=19.92%
T 1200

2. Between genes B and C = Q+R 128+124+9

------ x 100 = --------------x100=21.75%
T 1200

3. Between genes A and C = P+Q 230+252

------ x 100 = --------------x100= 40.20%
T 1200
Gene sequence

A 19.92cM B 21.75cM C

40.20cM

4. Combining map segments

Finally the different segments of maps of a complete chromosome are combined to form a
complete genetic map of 100 centiMorgan long for a chromosome.
Chromosome Map of Drosophila
The chromosome map of Drosophila includes four linkage groups corresponding to four
chromosome pairs. The genes present in the X chromosome constitute the first linkage group,
those present in 2nd and 3rd chromosome constitute 2nd and 3rd linkage groups and those on the
fourth chromosome form fourth linkage group. The fourth linkage group is the smallest of all.
Genetic map of Drosophila with four linkage groups

Chromosome maps of maize

Chromosome maps of maize have been drawn by R.A. Emerson. As there are 10 pairs
of chromosome 10 chromosome maps are seen.
Factors affecting the mapping
Chromosome map can be constructed only with the help of crossing over percentage.
The crossing over percentage is highly modified by the interference and coincidence.
Lecture 27. Sex determination: Autosomes and sex chromosomes - chromosomal
theory of sex determination- different types – sex determination in human, fowl,
butterfly, grasshopper, honey bee, fumea; Sex determination in plants –
Melandrium, papaya, maize.

Sex determination : It is the process of sex differentiation, which utilizes various genetic
mechanisms to decide whether a particular genotype will develop into male or female sex.
Sexual differentiation: The development of secondary sexual characters.
Primary sexual characters: The characters which are distinct since birth and take part in
reproduction such as the reproductive organs or gonads that produce gametes in
mammals.
Secondary sexual characters: Characters which express in adulthood differently in
males and females and do not take part in reproduction.
Sex chromosomes or allosomes: The chromosomes which differ in number and
morphology in male and female sex.
Autosomes: Those chromosomes which do not differ in number and morphology in male
and female sex.
Homogametic sex: Sex with similar sex chromosomes such as XX and ZZ and the
individual producing only one type of gamete with respect to sex chromosomes.
Heterogametic sex: Sex with dissimilar sex chromosomes such as XY, XO and ZW and the
organism will produce different type of gametes.

Sex determination is the process of sex differentiation, which utilizes various

genetic mechanisms to decide whether a particular individual will develop into male or
female sex. The reproductive organs (gonads) that produce gametes form the primary
sexual characters. The development of gonads in the body is the main character for sex
determination.
Other somatic differences of the male and female animals are called as secondary
sexual characters. The development of secondary sexual characters is called sexual
differentiation. Sex determination is by sex chromosomes and sexual differentiation is by
hormones.
SEX CHROMOSOMES: Sex determination in human beings and several other animals is
done with the help of specific chromosomes. This pair of chromosomes is called sex
chromosomes or allosomes, while the rest of the chromosomes are called as autosomes.
Sex chromosomes were first discovered by Mc Lung (1902) in grass hoppers and later by
Wilson and Stevens (1905) in Protenor. Diploid males have 13 chromosomes and diploid
females have 14 chromosomes. Therefore in pairing there will be one extra chromosome
in the male. This extra chromosome in males is known as the X-chromosome.
Structure of sex chromosomes
X chromosomes: In most of the organisms it is straight, rod like and comparatively larger
than Y chromosomes. It has large amount of euchromatin and small amount of
heterochromatin.
Y chromosome: It is smaller in size with one end slightly curved or bent to one side in
Drosophila ; in Man and Melandrium no such curvature occurs. It contains small amount
of euchromatin and large amount of heterochromatin. It has little genetic information,
referred as genetically inactive or inert.
Difference between autosomes and allosomes

SEX DETERMINATION AND SEXUAL DIFFERENTIATION IN HUMANBEINGS

Sex determination: Sex determination in human beings is by XY method, where
allosomes are X and Y chromosomes. The number of chromosomes in human being is 46.
Females are homogametic, AAXX and males are heterogametic, AAXY. The Y-chromosome
determines maleness in human beings.
The following syndromes are produced due to non-disjunction of allosomes in
human beings. These syndromes explain the importance of Y chromosome in the
determination of male sex in human beings.
1. Turner syndrome (2n=45 -44+X): In Turner syndrome, the affected individual
has female external genitalia and internal genital ducts, but the ovaries are rudimentary.
Other abnormalities are short stature, webbed neck and a broad, shield-like chest. It is due
to the monosomy of 23rd pair. Turner female is negative for Barr body.
2. Klinefelter syndrome (2n=47, 44XXY) : These individuals have male genitalia
and internal genital ducts, but their testes are underdeveloped. Feminine sexual
development is not completely suppressed. It is due to the trisomy of 23rd pair. Klinefelter
male is positive for Barr body.
Mechanisms of sex determination
1. Genic sex determination
2. Chromosomal sex determination
3. Environmental sex determination
4. Hormonal sex determination

1. Genic sex determination

Sex determination in plant species, e.g .papaya (Carica papaya), maize is governed
by genes. Genic sex determination is known in both monoecious and dioecious plants.

1.1. Sex determination in Dioecious plants

i. Dioecious plants produce flowers of only one gender or sex, and have male and
female [Link] breeding populations of many species are gynodioecious, consist
of plants having hermaphrodite flowers and those having female flowers.
ii. Some rare species are androdioecious as they have male and hermaphrodite flower
bearing plants.
iii. In these species, plants of different genders have different genotypes, and the
gender determining genes have the major effect on sex although environmental
factors can also affect the sex of flowers.
Sex determination in Papaya
In papaya, expression of sex is influenced "by a single gene” with three alleles m,
M1, M2.

Genotype Survival Sex-expression

mm Vital Female
M1m Vital Male

M2m Vital Hermaphrodite or bisexual

M1M1,M2M2 Lethal (all die) Sterile
and M1M2
 Genic sex determination was also reported in Spinach, (Spinacea oleracea)
Vitis cinerea and Asparagus.

1.2. Sex differentiation in monoecious plants

i. In monoeicous species, development regulation leads to production of flowers of
different genders. In maize and cucumber (cucumis sativus) several genes affect the
gender of flowers; mutations in these genes lead to feminization or masculinization
of flowers or to a change in the positions of male and female flowers.
ii. Generally, hormones affect flower sex, but the effects are species- specific.
iii. For example, ethylene and auxin application to cucumber flower meristems is
feminizing, while GA3, is masculinizing in contrast GA3 is feminizing in maize.
iv. In maize, gibberellins has feminizing action and gibberellin 1(GA1) is the major
biologically active gibberellin.

Sex determination in Maize

i. Maize plants are generally monoecious, A single recessive gene, ba for barren
plant in homozygous condition (baba) interferes in the cob development and
making functionally male
ii. Another recessive gene, ts for tassel seed in homozygous condition (tsts) converts
the male flowers into female flowers and plants do not produce pollen grains.
iii. In plants homozygous for both ba and ts are functionally female. So, the two
recessive genes (ba and ts) have converted a naturally monoecious plant into a
dioecious one.

Genotype Female flower Male flower Sex expression

BaBa TsTs Normal Normal Monoecious

baba TsTs Rudimentary Normal Male

BaBa tsts Normal Develop into Female

female flower
baba tsts Normal Rudimentary Female

In cucumber also, several genes affect the gender of flowers; lead to feminization or
masculinization of flowers or to a change in the positions of male and female flowers.
 Hormones also affect flower sex, but the effects are species- specific. For example,
ethylene and auxin application to cucumber flower meristems is feminizing, while GA3,
is masculinizing in contrast GA3 is feminizing in maize. Monogenic sex determination
also found in Chlamydomonas reinhardtii, Saccharomyces cerevisiae and Neurospora
crassa.

2. Chromosomal sex determination

It is grouped into three types
2.1. Theory of heterogamesis
2.2. Haplo-diploid mechanism – Honey bee
2.3. Genic balance theory - Drosophila
2.1. Theory of heterogamesis
This theory was proposed by Correns in 1906. In this method of sex determination
chromosomal number differs in male and female and one sex produces two types of
gametes.
There are four different systems of sex determination, viz.,
(1) XX - XY female-male system Heterogametic male system
(2) XX - XO female-male system
(3) XO - XX female-male system
(4) ZW - ZZ female-male system Heterogametic female system

Mechanism of sex Animals Plants

determination
(i) XX-XY System Drosophila, Human beings Melandrium, Cannabis,
and some other mammals Coccinia, Humulus lupulus, etc.
(ii) XX-XO System Grasshoppers, many Dioscorea sinuata (five leaf yam),
insects of Hemiptera and Vallisneria spiralis
Orthoptera. (common aquarium plant)
(iii) XO-XX System Fumea (Moth) —

(iv) ZW-ZZ System Birds, butterflies, Moths Fragaria (wild strawberry)

(XY-XX System)

A. Heterogametic male system

1. XX-XY method of sex determination

In human beings and insects like Drosophila both females and males have the same
number of chromosomes. Females are homogametic with XX chromosomes producing
similar ova having X chromosome. Males are heterogametic with X and Y chromosomes
producing two kinds of sperms, half of them with X chromosome and the other half with Y
chromosome. On fertilization the zygotes may have either the XX or XY. The zygote with
XX becomes the female and the zygote with XY becomes the male.
In human beings females are heterogametic with 44 autosomes and XX allosomes.
Males are heterogametic with- 44 autosomes and XY allosomes.
All the eggs are similar in their karyotype having 23 chromosomes, (22+X). Sperms
also have 23 chromosomes but are of two types. Half of them are with allosome X (22+X)
and the other half with allosome Y (22+Y). The sex of the offspring depends on the
fertilizing sperm.
In Melandrium album, the Y chromosome has 3 distinct regions influencing sex
determination and male fertility has been localized on the differential part of the Y
chromosome. The region I suppress the maleness. In the absence of this region, plants are
bisexual. Region II promotes male development. When this region (with or without Reg. I)
is missing, a female plant is produced. Region III carries male fertility genes, loss of this
region result in male sterility.

ii. XX-XO method of sex determination: In the insects of Hemiptera (bugs) and in
grasshoppers and many Orthoptera, females have two X chromosomes and males have
one X chromosome. The unpaired X chromosome determines the male sex. So females are
homogametic and males are heterogametic. The sex of the offspring depends on the
fertilizing sperm.
B. Heterogametic female: Two types
i. ZO-ZZ method of sex determination: In Fumea (moth), the female is heterogametic
with one Z chromosome (ZO) and male is homogametic with two Z chromosomes (ZZ). In
this “ZO” method of sex determination, the unpaired Z chromosome in the female
determines the sex. All the sperms are similar in their karyotype but ova differ in their
karyotype. The sex of the offspring depends on the fertilizing ovum.
ii. ZW-ZZ method of sex determination : In birds, reptiles, some fishes, butterflies etc.,
females are heterogametic with ZW chromosomes and males are homogametic with ZZ
chromosomes. All sperms are similar with allosome ‘Z’, but ova are of two different kinds.
Half of the ova are with allosome ‘Z and other half with allosome ‘W. The sex of the
offspring depends on the fertilizing egg.

2.2. HAPLO-DIPLOIDY IN HONEYBEES: In insects belonging to the order Hymenoptera

(honeybees), sex is determined by the number of sets of chromosomes (haploid or
diploid) a bee receives.

The eggs fertilized are diploid and the

unfertilized eggs are haploid (Apis mellifica
has 16 chromosomes in haploid state and
32 chromosomes in diploid state). Diploid
zygotes develop into diploid females.
Female that feeds on royal jelly become
the fertile female called queen and
others feed on honey become the sterile
females called workers. The unfertilized
eggs undergo parthenogenesis, which is
an asexual process of embryonic
development, to produce haploid fertile
males called drones. This parthenogenetic
development of haploid males from the
unfertilized eggs is called arrhenotoky.
Males produce sperms by mitosis rather
than meiosis.
Lecture 28- Genic balance theory of Bridges, quantitative theory, metabolic
differentiation theory, hormonal theory, barr bodies; Gynandromorphs – sex
reversal in chicken

Sex index : It is the ratio of X chromosomes to haploid set of autosomes.

Sex mosaic: Combination of male and female features in the body of an individual.
Gynandromorphs: Individuals with sex mosaic; also called as gynanders.
Barr body or sex chromatin: Relatively coiled and inactive chromatin of the X
chromosomes of the normal mammalian females but not the males.
Lyon hypothesis : This hypothesis states that an individual may have any number of X
chromosomes, but only one remains active and others become inactive or condensed.

2.3. GENIC BALANCE THEORY

This theory was formulated by Calvin Bridges. The sex determination in Drosophila is XY
method. The allosomes are X and Y chromosomes. Males and females have the same
number of chromosomes i.e. 8. Of these, six are autosomes and two are allosomes. Female
is homogametic AAXX and male is heterogametic AA XY, where A is the single set of
autosomes. He showed that Y chromosome has no role in the determination of sex in
Drosophila. He proposed that both the X chromosomes and autosomes together play a
critical role in the determination of sex in Drosophila.
Genetic balance theory states that sex determining genes are present in X chromosomes as
well as autosomes; male sex determining genes in autosomes and female sex determining
genes in X chromosomes. The sex expression is determined by the balance of genes on
autosomes and X chromosomes (or)
The expression of sex depends on the ratio of X chromosomes to haploid set of
autosomes; represented as sex index (X/A ratio).

He studied the offspring resulting from the non-disjunction of X chromosomes

during meiosis in females. Non-disjunction is the failure of the paired chromosomes to
segregate or separate during the anaphase stage of the first or second meiotic divisions of
gametogenesis.
The result is the production of abnormal gametes. One of which contains one extra
chromosome (AXX) and other contains one chromosome less (A). Fertilization of such
gametes with normal gametes produces aneuploid zygotes (2n+1 or 2n–1).
Bridges crossed a triploid female (AAAXXX) to a normal male and observed many
combinations of autosomes and sex chromosomes in the offspring. Triploid female
produces gametes viz., ‘AAXX’, ‘AX’, ‘AAX’, and ‘AXX’. The diploid male produces two types
of sperms viz., AX and AY. These female gametes fertilize with normal sperms of the male
producing different genotypes. From these karyotypes, Bridges found that the XXY flies
were normal females, and the XO flies were sterile males.

Gametes Xn (A)(AX) Yn (A)(AY)

X2n XX3n = 2X/3= 0.66 XY3n = 1X/3= 0.33

(AAX) Inter sex Super male or Metamale

Xn XX2n = 2X/2= 1.00 XY2n =1X/2= 0.50

(AX) Female Male
XX2n XXX3n = 3X/3= 1.00 XXY3n = 2X/3= 0.66 =
(AAXX) Triploid female Inter sex

XXn XXX2n = 3X/2 = 1.50 XXY2n = 2X/2=1.00

(AXX) Super female or Female
Metafemale

The presence of Y chromosome in the XXY flies did not cause maleness, and its
absence in the XO flies did not produce femaleness. He concluded that Y chromosome in
Drosophila lacks male-determining factors, but contains gametic information essential to
male fertility as the XO males were sterile. He realised that the factor in determining the
sex in Drosophila is the ratio of X
chromosomes to the number of haploid sets
of autosomes.
Normal females have a ratio equal to
1.0. If the ratio is >1.0, females are
metafemales, which are infertile. Normal
males have a ratio equal to 0.5. When the
ratio decreases the males are metamales, which are infertile. If the ratio is between 0.5
and 1.0, the flies are intersexes, which are larger, with morphological abnormalities and
rudimentary bisexual gonads. This mode of sex determination is explained as the genic
balance theory of Bridges. This theory explains that genes for maleness are located on
autosomes and for femaleness on X-chromosomes in Drosophila.

3. EFFECT OF ENVIRONMENT ON SEX DETERMINATION:

Specific sex determining genes and chromosomes are responsible for sex phenotype. The
genes for both maleness/femaleness are present in every species, and external factors
trigger the expression of either of the genes.
In Bonellia, sex determination is non-genetic and depends on environmental effect.
Bonellia is a marine, sedentary worm. Male and female have similar genotype, but
stimuli from the environment initiate the development towards one sex or other. The
female is big and it has a long proboscis. The males are small and lives as parasite in the
uterus of female. The larva produced by the female is hermaphrodites. The larvae move
away from the proboscis of female (the females of same species) grow into female. Larva
goes near the proboscis of any female of the species becomes male. The secretions of
proboscis in the female influence the young worms to develop in to males.
In Dinophilus, sex determination depends on size of the egg. Larger eggs after
fertilization develop into females and smaller eggs into males. Horsetail plant
(Equisetum), sex development depends on growing condition. Good condition-develop as
females and stress condition develop into males.
Cucumber and musk melon-treatment with ethylene, enhance production of
female flowers.
4. HORMONAL CONTROL OF SEX DETERMINATION:
Hormones are the secretion of the endocrine glands which are more prominent in
the sex differentiation rather than sex determination. Crew had reported a case of
complete sex reversal in fowls. A fertile hen may change to fertile cock due to the
damage to the ovary or after the natural cessation of egg-laying. The ovary in the
reproductive female secretes a male suppressing hormone. Hence male development is
suppressed as long as ovary is active. Testis may develop in the absence of the ovary or in
the natural cessation of ovary.
In cattle when fraternal twins, or dizygotic twins (one male and the other female)
are produced the female becomes sterile and the male becomes normal. This sterile female
co-twin is called freemartin. Male hormones are produced earlier. These male hormones
circulate to the female foetus during the development suppressing the development of
ovary in the female. So, the female co-twin becomes sterile.

Dosage Compensation
Human females (and females of all other mammal species) inherit two copies of
every gene on the X chromosome, whereas males inherit only one.
How is this difference in gene dosage compensated for? Or How to create equal amount of
X chromosome gene products in males and females?
To compensate the chromosome could do one of two things :
i. Double the amount of transcription of X-chromosome genes in males.
ii. Inactivate one of the X-chromosomes in females.
Females have only a single active X chromosome in each cell. The inactivation of one of
the two X chromosomes in female equalizes the gene dosage of X-linked genes. This is
known as dosage compensation. The inactivated X-chromosomes in females forms Barr
Bodies.

Barr bodies : M. L. Barr in cats, Keith Moore and [Link] in human being studied the
genetic dosage difference. Barr and Bertram observed darkly staining body in the
interphase nerve cell of female cats, but absent in male cats. This highly condensed
structure is called sex chromatin body or Barr body. Barr body is always one less than
the number of X chromosomes.
No. Barr bodies = N-1 (N = number of X chromosomes present)
46, XX 1 Barr body
45, X 0 Barr body
47, XXY 1 Barr body
47, XXX 2 Barr bodies
48, XXXX 3 Barr bodies
X-inactivation is the process of bringing equivalence in the expression of X-linked
genes in females and males. This inactivation event is irreversible during the lifetime of
the cell. X-inactivation can also occur in males with Klinefelter syndrome who have two X
chromosomes. Barr bodies are useful tools for detection of sex abnormalities and
determination of sex in humans. Individuals with Barr bodies are called as sex
chromatin positive and those do not have Barr bodies are sex chromatin negative.
Lyon hypothesis and dosage compensation
Mary Lyon suggested that this Barr body represents an inactive X chromosome. Barr body
is a tightly coiled heterochromatin. Lyon proposed that inactivation occurs randomly in
somatic cells at a point early in embryonic development. Once inactivation occurs, all the
progeny cells have the same inactivated X chromosome.
Sex mosaics or Gynandromorphs:
A gynandromorph is an organism which contains both male and female characteristics.
The term gynandromorph, from Greek words "gyne" female and "andro" male. These
characteristics can be seen in butterflies, where both male and female characteristics can
be seen physically because of sexual dimorphism. In Drosophila, some parts are female
and other parts are male. A clear example in birds is the gynandromorphic zebra finch. It
helps in the study of pathways of development, especially cell lineage.

Types of Gynandromorphs
1. Bilateral Gynanders
One half of the body shows female characters while other half shows male characters.
2. Anterior-Posterior Gynanders
In such gynanders, anterior region of the animal body has the characteristics of one sex
and posterior half region has the characteristics of the other sex. Eg. Bracon hebetor
(wasps)
3. Sex Piebalds
In these gynandromorphs, the body consists of female tissue having spots of male
tissue scattered irregularly. There are certain cases in which a few cells of the body show
sex difference.
Gynandromorphs are believed to arise from XX zygotes. Bilateral Gynanders arise
due to irregularity in mitosis at the first clevage of zygote (Mitotic non-disjunction). Thus
one daughter nucleus receives only one X chromosome, while other receives three X
chromosomes. During embryonic development, in one or more cells, one of the two X
chromosomes does not pass to any pole at anaphase and as a result, is lost. Thus one
daughter nucleus receives only one X chromosome, other receives two X chromosomes. If
loss occurs at later cell division, smaller proportion of adult body would be male. The
position and size of mosaic sector determined by place and time of division abnormality.
Lecture 29- Sex linked inheritance – criss cross inheritance – reciprocal difference;
holandric genes; sex influenced and sex limited inheritance inheritance-genetic
disorders.
Criss-cross inheritance: Inheritance of sex linked gens from grandfather to grandson
through daughter.
Sex linked traits or X linked traits: Characters for which the genes are located on X
chromosomes.
Sex linkage: The linkage of genes which are located on sex or X chromosome.
Sex limited traits: Characters which express in one sex only.
Sex influenced genes: Genes whose expression depends on the sex of an individual such
as baldness in human beings.

Sex linked inheritance

Inheritance of characters through X chromosome or on the analogous Z

chromosome is called sex linked inheritance. It was discovered by T.H. Morgan in
1910. Eye colour and bar eye in Drosophila, colour blindness and haemophilia in human
and barred plumage in fowls are inherited through X chromosome. Genes other than sex
determiners also located on the X chromosomes.

Characteristic features of sex linked genes

 Sex linked genes are located on ‘X’ chromosome only.

 In diploid, homogametic sex contains two copies of sex linked alleles where as
heterogametic sex contains only one sex linked allele.
 A recessive gene in a homogametic sex can express only when it is homozygous
state, whereas in heterogametic sex a recessive allele express in hemizygous
condition.
 Sex linked genes follow the criss cross inheritance.
 Sex linked gene exhibit several deviations from the normal segregation pattern.
Criss-cross inheritance of X-chromosome in Drosophila

Female is produced when an X egg is fertilized by X sperm. Male is produced

when Y sperm fertilizes an X egg.
 P Female Male

XX XY

Gametes (X) (X) (Y)

Progeny XX XY
A male receives an X-chromosome only from the mother and never from father.
The male receives Y chromosome only from his father, never front his mother. Thus
the inheritance of X chromosome in Drosophila follows specific pattern. The male
transmits his X chromosome to grandson only through his daughter. This is called
criss- cross inheritance. The female transmits the X-chromosome both to her son and
daughter. The criss – cross pattern of inheritance is characteristics of sex-linked
genes. This distinctive criss-cross pattern, from father through daughter to grandson
replacing the usual pattern for the F1 and F2 segregation is now interpreted as
evidence of sex-linkage.

Criss-cross inheritance of eye colour in Drosophila

A cross was made between white-eyed male Drosophila and red-eyed female
Drosophila by [Link] in 1910. The F1 flies were red eyed. When F1 files were
intercrossed, three fourth of the F2 flies possessed red eyes and one fourth white eyes.
From this familiar 3:1 ratio, it is clear that this is a monohybrid inheritance where red
is dominant over white. But, when the F2 flies were classified for both eye color and
sex, it was found that

[Link] the F2 females were red eyed.

[Link] of the F2 males were red eyed.
[Link] of the F2 males were white eyed.
When reciprocal cross was made between white eyed female and red eyed male
the F1 was composed of two different phenotypes ie., red eyed females and white
eyed males. When these F1 flies were intercrossed, the F2 consisted of flies in the
ratio of 1:1 for red eyed and white eyed. When these F2 files were classified for both
eye color and sex, it was found that
 i. Of two red-eyed flies, one is male and another is female.
ii. Of two white-eyed flies, one is male and another is female.
Reciprocal crosses for white-eyed Drosophila indicated that white-eyed is sex-linked

In the normal Mendelian inheritance, the F2 ratio does not differ from that of reciprocal
cross. But in the inheritance of eye color in Drosophila, the F2 ratio depends on the sex of
the parent by which eye color is introduced.
In Drosophila, the white eye colour follows a criss- cross inheritance. The kind of
inheritance from father to grandson only through daughter is called criss-cross
inheritance. The male transmits his red eye color to his grand sons through his daughters,
never to or through his sons. Thus, the transmissions of eye color and X chromosome are
similar. Hence, it is assumed that the gene for eye color is located in the X
chromosome and Y chromosome carries no allele for eye color.

Sex linkage in human beings

1. Colour blindness

The individual with colour blindness can not differentiate between red and
green colour. Colour blindness is recessive to normal.
 Daughters are normal in both cases, but they are the carriers of this trait

2. Haemophilia

Haemophilic individuals lack normal clotting of blood after its exposure to the air.
Normal – clotting 2 to 8 minutes after bleeding. Even small cut may prolong the
bleeding -leading to death. Hemophilia is a hereditary defect – governed by recessive
gene & inherited through females. The gene is located on X chromosome. In case of
marriage between haemophilic woman and normal man the disease will be
transmitted to 50% of the sons even if the gene is heterozygous condition.
Holandric genes

Most sex-linked genes in male heterogametic animals are on the X chromosome.

However, Y chromosome also contains few genes that produce visible effects on the
phenotype of the organism. Such genes are called Y linked or holandric genes.

X and Y chromosome contain some homologous segments. Genes on this segments are
incompletely sex linked (crossing over). Genes on non-homologous segment of X
chromosome are completely sex linked inheritance. Holandric genes reside in non-
homologous segment of Y chromosome. Holandric genes would be transmitted directly
from father to son and never appear in [Link] genes are in hemizygous state.

Characters controlled by holandric genes

1. Testis determining factor (TDF)

2. Hypertrichosis (excessive development of hairs on pinna of ear)
3. Genes for H-Y antigen (Histocompatibility antigen)
4. Spermatogenesis
5. Height and slower maturation of individuals

XY linked genes : XY linked genes are partially or incompletely sex linked because
sometime the cross over may occur in the homologous sections of X and Y linked
chromosomes. Eg. total colour blindness, skin diseases, Retinitis pigmentosa.

Sex-limited characters

Sex – limited genes are those which produce characteristics that are expressed in only
one of the sexes. Sex limited genes may be located in sex chromosome/autosomes. The sex
hormone is found to be limiting factor in the expression of sex limited gene. Sex limited
genes are responsible for secondary sexual characteristics.

For example beard in man and breast in women are produced by sex-limited genes.
A woman does not have a beard, though she carries all the genes necessary for beard.
Similarly man does not have breasts though he carries all the genes necessary for breast.
The expression of sex-limited characteristics depends upon the presence or absence of sex
hormones. Other examples include egg production in chickens; milk production in
mammals.

In humans
– Let B = beard and b = no beard.
– For males, BB and Bb have beards, while bb does not.
– For females, BB, Bb, and bb do not develop beards.
– Dominant gene B is expressed only in males.
Reason for difference between sexes is linked to sex hormones.

Chicken - cock feathering : In chicken the recessive gene (h) for cock feathering is
male sex limited.

Genotype Male Female

HH Hen feathering Hen feathering

Hh Hen feathering Hen feathering

hh Cock feathering Hen feathering

Sex influenced character

The condition in which the same gene acts as dominant in one sex and recessive in
other sex is called as sex influenced dominance. That is, the sex influences the gene
either to be dominant or recessive. The sex influenced genes are present in autosomes.
This differential behaviour of the gene is due to female and male sex hormones.
Expression of some genes is reversed depends on the sex of individual due to the internal
hormonal environment.
 For example, in human being baldness is due to sex influenced gene. This trait is
dominant in men and recessive in women. A man is bald in homozygous recessive as
well on heterozygous condition for baldness. Whereas women exhibit baldness only
in homozygous recessive condition for baldness and heterozygous condition for
baldness in female sex produce normal phenotype.

– Let bb = bald; b+b+ = not bald.

– b - acts like a dominant allele in males but recessive allele in females.
– For males bb and bb+ are bald, while b+b+ is not bald.
– For females bb is bald, while bb+ and b+b+ are not bald.
Lecture 30: Cytoplasmic inheritance and maternal effects – features of cytoplasmic
inheritance, chloroplast, mitochondrial - plastid colour in Mirabilis jalapa -
cytoplasmic male sterility in maize, kappa particles of paramecium

Non-Mendelian Inheritance

Maternal effects Cytoplasmic inheritance

Inheritance due to
Expression of Inheritance due to
organelle DNA
character influenced infective particles Plastids –Chloroplast
by the genotype of Inheritance associated
Mirabilis jalapa – four ‘O’ clock
the female parent with infective particles like
plant, Maize Iojab gene
parasite, symbiont or
Eg. Coiling pattern Mitochondria - CMS
viruses present in
of shell in snail
cytoplasm of an organism
Eg. Kappa particles in
Paramecium

Cytoplasmic inheritance

Besides chromosomes, various organelles of cytoplasm also contain DNA. The

mitochondria and plastids have their own DNA and carry their genetic characters
themselves. The mechanism in which cytoplasmic inclusions (e.g., alpha, beta, sigma and
kappa particles) and organelles (plastids, mitochondria, centriole, etc) take part in
transmission of characters from generation to generation is called as cytoplasmic
inheritance. Since cytoplasmic inheritance is based on cytoplasmic DNA molecules, it is
also called extra chromosomal inheritance.

Genes located in the cell organelle in the cytoplasm are termed as cytoplasmic genes or
plasma genes or cytogenes or extra nuclear genes or extra chromosomal genes and all the
plasmagenes of a cell constitute the Plasmon (like the genome).Cytoplasmic inheritance is
due to the plasmagenes located in cell organelles that are integral constituents of normal
cells. The plasma genes are capable of self duplication and mutation
Characteristics of cytoplasmic inheritance

1. Different in reciprocal crosses

In Mendelian inheritance, the results of reciprocal crosses are identical (one exceptional –
sex linked inheritance). If the character is transmitted through cytoplasm, the reciprocal
cross results will be different.
2. Somatic segregation
Plasma genes generally show somatic segregation during mitosis, a feature of rare
occurrence in the case of nuclear genes.
3. Non-mappability
Gene controlled characters shows linkages and hence they are mappable. But the
characters transmitted through cytoplasm show no linkage. Hence, they are not mappable.
4. Non-Segregation
The cytoplasmic heredity fails to show segregation. Sometimes, segregation may occur in
cytoplasmic heredity also. But it will not be consistent with the segregation of
chromosomes.
5. Indifference to nuclear substitution
When the nucleus is transplanted, no change is found in the cytoplasmic inheritance.
6. Infection like transmission
Cytoplasmic inheritance seems like infection through some agents.

Differences between maternal effect and cytoplasmic inheritance

Maternal effect Cytoplasmic inhertance

Inheritance of a trait controlled by the Inheritance of a trait controlled by genes in

genotype of the mother the cytoplasm rather than nuclear genes

Inheritance due to nuclear genes Inheritance due to plasma genes

Involved the inheritance by the nuclear Involved the plasmagenes of the mother
genes of the mother

[Link] coiling in snails [Link] inheritance in 4’O clock plant

Differences between nuclear and cytoplasmic inheritance

Mendelian Inheritance Cytoplasmic inhertance

Nuclear genes Plasma genes

Distinct segregation pattern Does not exhibit segregation

No reciprocal difference Reciprocal difference observed

Does not show maternal effect Exhibit maternal effect

Genes can be easily mapped on Mapping of plasma genes difficult

chromosomes
Linkage relationship among nuclear No linkage relationship among cytoplasmic
genes genes

Nuclear genes associated with Chloroplast DNA or mitochondrial DNA

chromosomes

1. Maternal effects

Maternal effect refers to the inheritance pattern of nuclear genes in which the
genotype of the mother directly determines the phenotype of the offspring, regardless of
the genotype of the father or offspring. Maternal effects are controlled by nuclear genes of
the mother. Characters showing the maternal effect exhibit clear-cut differences in F1 for
reciprocal crosses.
Maternal effect was first described by Sturtevant in his studies on coiling pattern
of shell in water snail, Limnea peregra. In this snail, the direction of coiling of shell is
controlled by single nuclear gene D/d; the dominant allele D produces right-handed
coiling (Dextral), while its recessive allele d produces left-handed coiling (Sinistral). The
direction of shell coiling in an individual is governed by the genotype of its female parent
and not by its own genotype. As a result, reciprocal crosses show differences in coiling in
F1 and there is no phenotypic segregation in F2 the phenotypic effect of segregation is
observable in F3 only.
Crosses between females with left-handed coil (dd) and males having right handed
coil (DD) produce F1 progeny (Dd) with left-handed coil, since the genotype of the female
parent is dd. In F2 segregation of Dd produces three genotypes (DD,Dd,dd) in the ratio of
[Link]. But the F2 snails with DD, Dd as well as dd genotypes exhibit right handed coiling
since their female parent has the genotype Dd which determines right-handed coiling in
the progeny (irrespective of the genotypes of the progeny). The F3 progeny from the F2
individuals with the genotypes DD and Dd showed right handed coiling, while those from
dd F2 individuals exhibited left-handed coiling of their shells; thus produces the typical
3:1 ratio in F3.
The reciprocal cross (DD x dd), on the other hand, yields right-handed coiling in the F1
(Dd) as well as in the three genotypes, [Link], obtained in the F 2. But in F3 2/3 of
the progenies showed right-handed coiling since they were derived from F2 individuals
having the genotypes DD and Dd. The remaining 1/3 of the F3 progenies exhibit left
handed coiling since their female parents had the genotype dd; this yield the typical
monohybrid ratio of 3:1 in the F3.
Maternal Effects and Embryonic Development

The molecular basis of maternal effect can be explained by examining the process of
oogenesis in females. Surrounding the oocytes are the nurse cells (2n). In the nurse cells,
both copies of the gene encoding coil orientation are active. The nurse cells transport their
gene products (mRNA and proteins) into the oocyte. If the Dd or DD are released into the
oocyte, it will have dextral coiling. If dd is released into the oocyte, it will have sinistral
coiling. Direction of coiling is determined by the orientation of the mitotic spindle in the
first mitotic division after fertilization. Maternal products within the oocyte direct
orientation of the mitotic spindle, and thus shell coiling.
2. Cytoplasmic inheritance

2.1. Inheritance due to kappa particles in Paramecium

There are two types of strains in Paramecium. One has kappa particles in its cytoplasm
and other does not have such particles in its cytoplasm. The presence of kappa particles in
the cytoplasm leads to production of a toxin known as paramecin. This toxin can kill the
strain Paramecium that lacks kappa particle. Thus the strain with kappa particle is known
as killer strain and that without kappa particle is called as sensitive strain.

The production of kappa particles is dependent on a dominant allele K, so that the killer
strains are KK or Kk and sensitive strains are ordinally kk. In the absence of dominant
allele K, kappa particles can not multiply and in the absence of kappa particles, dominant
allele K cannot produce them de novo.

If the killer (KK) and sensitive (kk) strains are allowed to conjugate, all exconjugants (the
cells separating after conjugation) will have the same genotype Kk. The phenotypes of
these exconjugants will however depend upon duration for which conjugation is allowed.
If conjugation does not persist long enough for exchange of cytoplasm, heterozygote (Kk)
exconjugants will only have parental phenotypes. It means that killers will remain as
killers and sensitive will remain as sensitive after conjugation. If conjugation persists,
sensitive stain will receive kappa particles and will become killer, so that exconjugants
will be killers having genotype Kk.

2.2. Cytoplasmic inheritance through organelles

[Link] inheritance

Plastids are minute cytoplasmic organelles in plant [Link] of the plant

cell contain circular DNA molecule which are self – replicating in nature. They are
transmitted through the cytoplasm of the egg. The isolated chloroplasts are capable of
protein synthesis in the presence of light. The DNA analysis revealed that 30-60 copies of
the chloroplast genome are found in each chloroplast of higher plants. The chloroplast
genome contains herbicidal resistant and streptomycin resistant genes.
Shoot Variegation in four O’Clock

The Four-O’clock plant, mirabilis jalapa, has branches that produce either green, white or
mixed green-white (variegated) leaves. In crosses between flowers of these branches, the
offspring are all green if the maternal parent is a flower from a green branch. Such
offspring remain green throughout subsequent generations as long as maternal plant is
green. Similarly, as long as the maternal parent is from a white branch, the offspring are all
white. When variegated branches are used as female source, both green and plastids are
present in cells of female parent. Therefore, female gametes may carry either green or pale
plastids or both. Consequently, three kinds of plants namely green, pale and variegated
plants would be obtained.

Egg source Pollen Source Progeny

White Green White

White
Variegated
Green Green Green
White
Variegated
Variegated Green Green
White White
Variegated Variegated

2. Mitochondria (mt DNA)

Mitochondria are present in living organisms arise from pre existing mitochondria. They
are small cytoplasmic organelles present in animal and plant cells but not present in
bacteria and viruses. Mitochondria provide cellular energy through oxidative
phosphorylation. Mitochondria contain a small circular DNA molecule codes for limited
number of structures and functions. The size of mtDNA ranges from about 16 kb in
mammals upto several hundred kilo base pairs in higher plants (eg. 570 kb in maize) and
mt DNA usually found in multiple copies per organelle. The mtDNA play a significant role
in crop improvement. Recent evidences showed that the cytoplasmic genetic male sterility
system in crop plants is due to the interaction of mitochondrial genome to the nuclear
genome.
Cytoplasmic male sterility (CMS)

In case of male sterility, pollen grains of male sterile are aborted. This male sterility
is transmitted only through the female and never by the pollen. When all of the
chromosomes of the male sterile line were replaced with chromosomes of normal plants,
the line still remained male sterile, showing thereby that male sterility is controlled by
some agency in the cytoplasm. It was later recognized that cytoplasmic male sterility in
maize results from alterations in the heredity units in the mitochondria (mitochondrial
DNA). In several crops like rice, maize, sorghum, pearl millet cytoplasmic control of male
sterility is known. In such cases if female parent is male sterile, F1 progeny would always
be male sterile because cytoplasm is mainly derived from egg obtained from male sterile
female parent.

CMS in maize

Three types of CMS viz., CMS-T, CMS-C and CMS-S have been identified. Among this, CMS is
commonly used for the production of hybrids since it is highly stable under all
environmental conditions. It is characterized by failure of anther exsertion and pollen
abortion. Mitochondrial T-urf 13 is a unique sequence responsible for tapetum
degeneration during microsporogenesis. This in turn causes pollen abortin and finally
leads to male sterility

3. Plasmids

Plasmids are extra chromosomal, circular, covalently closed double stranded DNA
molecules found in bacteria. Plasmids can replicate autonomously of the host
chromosome. The size of plasmid ranges from two to several hundred kilobases. Plasmids
carry genes for the inactivation of antibiotics, metabolism of natural products and
production of toxins. The F factors and R factors are important plasmids of Escherichia
coli.
Plasmid Ti (tumour inducing) carries DNA sequence-transform cells of dicot plant
to tumour cells. Disease induced by the bacterium Agrobacterium [Link]
bacterium initiated the gall disease are killed and fragment of Ti Plasmid carried by the
bacterium combined with DNA fragment of infected plant. The DNA carried by the plasmid
integrated into plant cells. The plasmid integrated into chromosomal genome and stable in
inheritance and such plasmids are called as episomes.
Lecture 31- DNA, the genetic material – Griffith’s experiment, experiment of Avery,
McCleod and McCarthy – confirmation by Hershey and Chase; RNA as genetic material –
Frankel, Conrat and Singer experiment.
Linear DNA: DNA which has a thread like structure with both ends free. Such DNA is found
in Eukaryotes.
Circular DNA: DNA which has a ring or circular shape. Such DNA is usually found in
Prokaryotes, chloroplast and mitochondria.
Double stranded DNA: DNA which has spirally arranged double strands. It is found in all
plants, animals and bacteria.
Single stranded DNA: DNA with one helix.
Transformation: The genetic recombination in which naked DNA from one cell enter and
integrate into another cell.

[Link] as genetic material

DNA is a nucleic acid that carries the genetic information in cells consisting of two long chains
of nucleotides twisted into a double helix and joined by hydrogen bonds between the
complementary bases adenine and thymine or cytosine and guanine. DNA sequences are
replicated by the cell prior to cell division.

It was postulated by Mendel in 1865 that the development of characters in peas (and other
organisms) was controlled by particulate factors; later, these factors were later called genes. In
1902, Sutton and Boveri postulated that genes were located in chromosomes; today this is
universally accepted. However, the chemical nature of genes became evident through a series
of brilliant experiments during the 3rd to 6th decades of the last century.

Properties of Genetic Material

1. Repository of genetic information

2. Information must be transmissible to progeny with high precision
3. Physical and chemical stability
4. Potential for heritable change should be low
The process of identification of genetic material began in 1928 with the experiments of
Griffith and concluded in 1952 with the studies of Hershey and Chase. Another ingenious
 experiment carried out independently by Gierer and Schramm and Frankel-Conrat in 1956
established that in some viruses, RNA functions as the genetic material.

Experiments for DNA as genetic material

1. Griffith‟s experiment
2. Experiment of Avery, McLeod and McCarthy
3. Hershey and Chase experiment
1. Griffith experiment (1928)

The phenomenon of transformation in pneumococcus pneumoniae bacterium (Streptococcus

pneumoniae) was discovered by Fredrick Griffith in [Link] are two types of
pneumococcus bacteria – virulent (pathogenic) and avirulent (non-pathogenic). Virulent strains
have polysaccharide capsules and give smooth colonies (S). Avirulent strains have no capsules
and give rough colonies (R).

Virulence Colony morphology Designation

Virulent Smooth III S
Avirulent Rough II R

Live II R and heat killed IIIS are not lethal, when injected into the mice separately. But a
mixture of live II R and heat killed IIIS was lethal to the mice. The blood of dead mice
contained live IIIS. The heat killed IIIS had transformed live II R into live IIIS. Griffith
(1928) called this phenomenon as transformation. Thus the transformation of non virulent
Type II R cells to virulent Type IIIS cells cannot be explained by mutation, rather some
component of the dead Type IIIS cells must convert living Type II R cells to Type
[Link] hypothesized that the transforming agent was a “IIIS” protein.

The same phenomenon occurred in the test tube when live Type II R cells were grown in the
presence of dead type IIIS or extract of Type IIIS was hereditary and it set the stage for
determining the chemical basis of hereditary in Pneumococcus.

2. Experiment of Avery, Macleod and McCarty (1944)

The first direct evidence showing that the genetic material is DNA rather than protein or
RNA was published by O.T. Avery, C.M. Macleod and M. Mc Carty in 1944. The most definite
experiments conducted by them proved that the DNA was the transforming principle (DNA is
the genetic material) involved the use of enzymes that degrade DNA, RNA, or protein. In
separate experiments highly purified DNA from Type IIIS cells was treated with (1)
Deoxyribonuclease (DNase which degrades DNA) (2) Ribo nuclease (RNase which degrades
RNA) or (3) Proteases (which degrade proteins) and then tested for its ability to transform Type
II R cells to Type IIIS. Only Deoxyribonuclease had any effect on the transforming activity of
the DNA preparation. It totally eliminated all transforming activity.

The results obtained by Avery and co-workers clearly established that the genetic
information in pneumococcus was present in DNA. We now know that the segment of DNA in
the chromosome of pneumococcus that carries the genetic information specifying the synthesis
of a Type III capsules is physically integrated into the chromosome of the Type II R recipient
cell by a specific recombination process occurring transformation.

The findings of Avery and coworkers presented indisputable evidence for DNA to be the
transforming principle. But in spite of this, DNA could not be universally accepted as the
genetic material. One of the main reasons for this was the lack of information on the molecular
mechanism of transformation.

3. The Hershey-Chase Experiment (1952)

The additional direct evidence indicating that DNA is the genetic material was published in
1952 by A.D Hershey (1969 Nobel Prize Winner) and [Link]. These experiments showed
that the genetic information of a particular bacterial virus (bacteriophage T2) was present in
DNA. It was the universal acceptance of DNA as the genetic material.

Viruses are the smallest living organisms. They never as such enters the cell; only the
tail contacts the host and enzymatically cuts a small hole through the membrane and then the
nucleic acid of the virus head flows to the cell. This idea was tested by Hershey and Chase in
the following way. Phage DNA was labelled with radio-isotope 32P in place of normal isotope,
whereas protein coat was labelled with 35S in the place of normal isotope 32S. These labels are
highly specific, because DNA does not contain sulphur and the protein coat is devoid of
phosphorus. A sample of an [Link] culture was infected with labelled T2 phage. After a short
incubation period, the suspension was spun for a few minutes in warring Blender at 10,000
rpm. The resulting suspension was centrifuged. The pellet contained infected bacteria, where as
32 35
supernatant contained smaller particles. These fractions were analysed for P and S to
determine the location of the phage DNA and the protein coat. The results of the experiment
were:

 Most of the phage DNA was found in the bacteria.

 Most of the phage protein was found in the supernatant.
 The blender treatment did not prevent the infection.
 The progeny of T2 phage contained the parental 32P and not the parental 35S.
This led to the conclusion that the phage has a genetic part-DNA and non-genetic
protective part-protein. The protein coat serves as a vehicle. Only DNA carries necessary
information for the new generation of phages.

B. RNA as genetic material in small viruses

In viruses, the genetic information are present either in DNA or RNA. Tobacco Mosaic
Virus (TMV) is an RNA virus. It consists of a single molecule of RNA surrounded by a protein
coat. By using the appropriate chemical treatments, one can separate the protein coats of TMV
from RNA. Moreover, this process is reversible; by mixing the proteins and RNA under
appropriate conditions “reconstitution” will occur producing complete infective TMV particles.
Fraenkel-Conrat and Singer (1957) took two different strains of TMV, separated the RNA
from the protein coats and reconstituted mixed virus by mixing the proteins of one strain with
the RNA of second strain and vice-versa. When these mixed viruses were used to infect tobacco
leaves, the progeny viruses produced were always found to be phenotypically and genotypically
identical to the parent strain from which the RNA had been obtained. Thus it was proved that
the genetic information of TMV is stored in RNA and not in protein.
Lecture 32-Structure of DNA – Watson and Crick model Proof for semi conservative method
of DNA replication; Models of DNA replication; steps involved in DNA [Link] types
- mRNA, tRNA, rRNA – Protein synthesis

DNA : It is a double helical structure of two polynucleotide strands that are spirally twisted
around each other and coiled around a common axis to form a right-handed double-helix.
Right handed DNA: An usual form of DNA with coiling of the helix in the right direction.
Left handed DNA: DNA with coiling of the helix in the left direction such as Z-DNA.
Nucleoside:A combination of deoxyribose sugar and nitrogen base.
Nucleotide: It is the repeating subunits of each polynucleotide chain. Each nucleotide is
composed of i. a phosphate group; ii. a five carbon sugar (pentose) and iii. a nitrogenous base.
Purines: The nitrogenous bases with double ring structure. They are of two types viz., adenine
and guanine in DNA and RNA.
Pyridimines: The nitrogenous bases with single ring structure. They are of two types viz.,
cytosine and thymine in DNA and cytosine and uracil in RNA.
DNA replication: The process by which a DNA molecule makes its identical copies. It is of
three types, viz., dispersive, conservative and semi-conservative.
Dispersive replication: DNA replication in which the new DNA molecules have old and new
DNA in patch.
Conservative replication: DNA replication in which a new DNA molecule has parental strands
and other contains newly synthesized strands.
Semi-conservative replication: DNA replication in which each of two new DNA molecules
has one old and one new strand. This is universally accepted.
Leading strand:The continuously replicating strand of DNA molecule.
Lagging strand:The discontinuously replicating strand of DNA molecule.
Okazaki fragments:Short segments of nucleotides synthesized in lagging strand of DNA as a
result of discontinuous replication.
Template: A macro molecule which provides information for the synthesis of another
complementary macro molecule.
Sense strand: One of the two complementary strands of DNA used as template for the
synthesis of RNA.
Non-Sense strand: The DNA strand which is not used for the synthesis of RNA.
Bi-directional replication:Replication of DNA in both directions from the point of origin.
Messenger RNA: The RNA which carries information from nuclear DNA to cytoplasm for
protein synthesis.
Ribosomal RNA: RNA which is found in the ribosomes in the cytoplasm.
Transfer RNA: The RNA which carries amino acids and attaches them with ribosome mRNA
complex for use in protein synthesis. Also known as soluble RNA.
Genetic code: The relationship between the sequence of bases in RNA and the sequence of
amino acids in a polypeptide chain.
Codon:Triplet sequence of RNA bases which codes for a particular amino acid.
Sense codons: Those codons which code for amino acids.
Signal codons: Codons which code for either start or stop signal during protein synthesis.
Anti codons:The base sequence of tRNA which pairs with codon of mRNA during translation.
Inosine: A newly discovered nucleotide which is found in third position in a codon and can pair
with A, U and C.
Wobble base pairing: The pairing of mRNA codons with tRNA anticodon in which first two
bases have normal pairing and the third base has abnormal pairing.
Protein: A polymer of amino acids.
Polypeptide: A product of the union of several amino acids.
Transcription: The process of synthesis of messenger RNA from a DNA template.
Translation: The process of protein synthesis from the information in mRNA.

Structure of DNA molecule (Watson and Crick’s DNA double helix model)
The term DNA was given by Zaccharis. In 1953, J.D. Watson (an American biologist) and
F.H.C. Crick (a British Physicist) proposed the three-dimensional model of physiological DNA
(i.e B-DNA) on the basis of X-ray diffraction data of DNA obtained by Franklin and Wilkins.
For this epoch-making discovery, Watson, Crick and Wilkins got Nobel Prize in medicine in
1962. The double helix model proposed by them is based on two evidences.

1. Chargaff’s chemical analysis

Chargaff (1950) found that a specific quantitative relationship is present between purines and

A+G = T+C
A = T
G = C
AT = GC
 pyrimidines of DNA molecule. In DNA molecule, the ratio of adenine to thymine and guanine
to cytosine are 1:1 that is amount of purine = amount of pyrimidine. It is called as Chargaff rule.

2. Crystallographic studies by Wilkins and Franklins

The X-ray diffraction patterns and crystallographic

data on DNA structure from the studies of
M.H.F. Wilkins and R. Franklins showed that DNA
is highly ordered multiple strand structure with
repeating sub unit structures spaced in every 3.4 Å
along the axis of the molecule.

Structure of DNA

On the basis of Chargaff‟s chemical data and

crystallographic data by Wilkins and Franklins, Watson and Crick proposed the structure of
DNA. The important features of their model are

i. The DNA molecule consists of two polynucleotide strands that are spirally twisted around
each other and coiled around a common axis to form a right-handed double-helix.
ii. The two strands are anti-parallel i.e. they run in opposite directions so that the 3′ end of one
chain facing the 5′ end of the other. This mechanism is very important in considering the
mechanism of replication of DNA.
iii. Each polynucleotide chain consists of sequence of nucleotides linked together by
phosphodiester bonds and two polynucleotide chains held together by hydrogen bonding
between bases.
iv. The purine and pyrimidine bases are on the inside of the helix where as the phosphate and
deoxyribose unit are on the outside.
v. The base pairs are stacked between two chains perpendicular to the axis of the molecule
similar to the steps of a spiral staircase. The base pairs in DNA stacked 3.4 Å apart with 10
base pairs per turn (34 Å) of the double helix.
vi. The base pairing in DNA molecule is specific ie Adenine pairs with Thymine (A=T) by two
hydrogen bonds and Cytosine pair with Guanine (C=G) by three hydrogen bonds. So each
base pair consists of one purine and one pyrimidine. This complementarity is known as the
base pairing rule.
vii. The high degree of stability of DNA is due to more number of hydrogen bonds.
viii. The DNA helix has a shallow groove called minor groove (1.2nm) and a deep groove called
major groove (2.2nm) across.
Each nucleotide in DNA is made of

 Deoxyribose sugar

 a phosphate group

 one of four nitrogen bases

Nitrogenous base + deoxyribose =
deoxyribonucleoside
Deoxyribonucleoside + Phosphate group =
deoxyribonucleotide

Conformations of the DNA double helix

The most common conformation in most living cells (the one depicted in most diagrams of
the double helix, and the one) is known as B-DNA proposed by Watson and Crick.
There are also two other conformations or Forms:
A-DNA, a shorter and wider form that has been found in dehydrated samples of DNA and
rarely under normal physiological circumstances
Z-DNA, a left-handed conformation. It is a transient form of DNA, only occasionally
existing in response to certain types of biological activity. Scientists have since discovered
that certain proteins bind very strongly to Z-DNA, suggesting that Z-DNA plays an
important biological role in protection against viral disease (Rich & Zhang, 2003).

Comparison between DNA and RNA

Characters DNA RNA

Pentose Deoxyribose Ribose
Base Thymine present and uracil Uracil present and thymine absent.
absent
Number of strands double-stranded single-stranded
Function Genetic material only  Generally, function as
mRNA rRNA, tRNA
 In some viruses; it serves as
genetic material.
Origin Replication of pre Through replication of RNA by
existing DNA RNA dependent RNA ploymerase
Native form Double-stranded DNA Double-stranded RNA usually in A-
usually in B-form form
Central dogma of life

The central dogma of molecular biology describes the flow of genetic information in cells
from DNA to messenger RNA (mRNA) to protein. It states that genes specify the sequence of
mRNA molecules, which in turn specify the sequence of proteins.
Transcription is the process of creating a complementary RNA copy of a sequence of
DNA. Both RNA and DNA are nucleic acids, which use base pairs of nucleotides as a
complementary language that enzymes can convert back and forth from DNA to RNA. During
transcription, a DNA sequence is read by RNA polymerase, which produces a complementary,
antiparallel RNA strand.
Translation is the process by which mRNA is decoded and translated to produce a
polypeptide sequence, otherwise known as a protein. This method of synthesizing proteins is
directed by the mRNA and accomplished with the help of a ribosome, a large complex of
ribosomal RNAs (rRNAs) and tRNA.
DNA replication

Watson and Crick proposed that during DNA replication, the two strands of DNA molecule
separate after the breakage of hydrogen bonds and each strand acts as a template for the
synthesis of a new companion strand. Thus, resulting daughter DNA molecules each
containing an old strand derived from parent DNA molecule and another strand newly
synthesized. This type of distribution of parental strands is called as semi-conservative.
However in considering possible mechanisms of DNA replication, three different hypothetical
modes are apparent in addition to semi conservative method of replication.

1. Semi – conservative method

In this method, two strands of the parental DNA molecule separates and each strand act as a
template for the synthesis of new complementary strand. Thus resulting daughter DNA
molecules each contain one old strand derived from parental DNA molecule and another strand
newly synthesized.

2. Conservative method

In this method, two parental DNA strand separates and each strand act as template for
synthesizing a complementary strand. The resulting progeny DNA molecule composed of two
newly synthesized strand and parental DNA strands remain intact (totally conserved after
replication)
3. Dispersive method

In this method a segments of parental strands and progeny or nascent strands become
interspersed through some kind of fragmentation, synthesis and rejoining process.

Semi Conservative Replication

The semi conservative mode of DNA replication was postulated by Watson and Crick
along with the double helix model of DNA. This method of DNA replication is universally
accepted because there are several evidences in support of semi conservative method.

Proof for semi-conservation method of DNA replication

The semi conservative mode of DNA replication was postulated by Watson and Crick
along with the double helix model of DNA. This method of DNA replication is universally
accepted because there are several evidences in support of semi conservative method. Various
experiments have demonstrated the semi-conservative mode of DNA replication. There are
three important experiments, which support that DNA replication is semi-conservative.

(1) Meselson and Stahl experiment

(2) Taylor‟s experiment
(3) Cairns experiment
Meselson and Stahl’s Experiment

The evidence for semi-conservative replication of DNA was first presented by Meselson and
Stahl in 1958. They grew [Link] on 15N (a heavy isotope of 14N) for 14 cell generations so that
the nitrogen present in DNA bases of these cells was 15N. DNA having 15N has a higher density
(1.724 g/cm3) than that having 14
N (1.710 g/cm3) therefore, they are called heavy and light
DNA. Heavy and light DNAs readily separated through cesium chloride density gradient
equilibrium [Link] form distinct bands in the centrifuge tube. In density gradient
centrifugation, a heavy salt solution, e.g.,6M CsCl2 is centrifuged at 30,000-50,000 rpm for 48-
72 hrs. This leads to the formation of a linear gradient of increasing density from the top to the
bottom of centrifuge tube.

14
Then these bacterial cells were transferred to N medium. The bacterial cells were sampled at
various times to ascertain the density of DNA. The sample time corresponded with the doubling
of the cells. After one generation, the DNA of daughter bacteria had neither original 15N density
nor the pure 14N density. Instead, this DNA had an intermediate (or) hybrid density (precisely
between 15N – 14N densities). The absence of 15N DNA indicated that the parental DNA was not
conserved as an intact unit on replication. The absence of 14N DNA indicated that the daughter
DNA molecule was not synthesized entirely denovo (afresh).Of two strands of the daughter
DNA molecule, one strand was
15
derived from the parent N
DNA and the other strand was
14
newly synthesized from N
source. Hence, the daughter
DNA molecule, being hybrid in
nature (15N14N) gives an
intermediate (or hybrid) density.
14
After two generations of bacteria in N medium there were equal amounts of hybrid DNA
15 14 14
( N N) and ordinary DNA ( N).

Meselson and Stahl concluded that during DNA duplications, each daughter molecule receives
one strand from the parent molecule. This strand is conserved through much duplication. Their
results agreed perfectly with the Watson-Crick hypothesis of DNA replication.
Features of DNA Replication

1. DNA replication is semi-conservative- Each strand of template DNA is being copied.

2. DNA replication is bidirectional-Bidirectional replication involves two replication forks,

which move in opposite directions.
3. DNA replication is semi-discontinuous- The leading strand copies continuously and the
lagging strand copies in segments (Okazaki fragments) which must be joined.
Steps in DNA replication

1. Opening DNA double helix

i. Initiation of Replication
The origin of replication is a unique DNA sequence at which DNA replication is initiated.
DNA replication may proceed from this point bidirectionally or unidirectionally. This origin of
replication is unwind, with one replication fork on either end. The replication fork is a
structure formed when DNA is ready to replicate itself.

ii. Unwinding of strands

The two stands of DNA double helix unwind and separate. Each strand serves as a template for
the synthesis of new strand. The unwinding takes places with the help of 3 types of protein.

[Link] unwinding protein or DNA helicases - unwinds and splits the DNA ahead of the fork.
b. Single strand binding protein (SSB)- Binds to single stranded region of DNA by the action
of helicase and thus preventing the strands from reuniting.
[Link] gyrase -Maintains proper topographical structure in the replication fork. This origin of
replication is unwind, with one replication fork on either end.

2. Building a primer
Synthesis of RNA primer is essential for initiation of DNA replication by DNA Polymerase III.
RNA primer is synthesized with the help of a special type of Primases. Primase is a form of
RNA polymerase. Primase is activated by DNA helicase. It synthesizes a short RNA primer
approximately 11 ±1 nucleotides long, to which new nucleotides can be added by DNA
polymerase III.
3. Assembling complementary strand

i. DNA synthesis starts on the RNA primer.

ii. A molecule of DNA polymerase III binds to one strand of the DNA. It moves along the
template strand, for assembling a leading strand (red) of nucleotides and reforming a
double helix.
iii. A second DNA polymerase III molecule (also green) is used to bind to the other
template strand. This molecule synthesizes discontinuous segments of polynucleotides
called as Okazaki fragments.
iv. The replication may take place either in one direction or in both the directions from the
point of origin.
4. Elongations of new DNA strand

i. DNA polymerase III begins to add deoxyribonucleotides to the 3‟ – hydroxyl end of the
RNA primer. The chain elongation reaction occurs by means of a nucleophilic attack of
the 3‟ – OH terminus of the primer on the inner most phosphorus atom (phosphorus atom)
of the incoming deoxyribonucleoside triphosphate. A phosphor diester bridge is formed
and pyrophosphate is released. The subsequent hydrolysis of pyrophosphate drives the
polymerization forward.

ii. The elongation of the DNA chain proceeds in the 5” 3” direction. The
polymerization is processive – that is, many nucleotides are added without the release of
the enzyme from the template. DNA polymerase catalyses the formation of a phosphor-
diester bond only if the base on the incoming nucleotide is complementary to the base on
the template strands. Thus DNA polymerase is a template directed strand.
5. Removing the primer
DNA polymerase I degrades the RNA primer.
6. Joining of Okazaki Fragments
The discontinuous fragment of Okazaki is joined to make continuous strands. The enzyme,
DNA ligase (violet), stitches the Okazaki fragments together into the lagging strand.

Ribonucleic acid

RNA is found in cells of all living organisms. It contains ribose sugar, nitrogen bases
and phosphate group. The nitrogen bases include adenine, guanine, cytosine and uracil and
pairing occurs between AU and GC. The function of RNA is transfer of genetic message from
nucleus to the cytoplasm and synthesis of protein in the ribosomes. In some viruses, RNA acts
as the genetic material and regulates the gene action.

Flow of genetic information

DNA RNA Protein

(Transcription) (Translation)

Structure of RNA

RNA is a long unbranched polymer consists of nucleotides joined by phosphor-diester

bonds. RNA differs from DNA in the following ways.

 RNA is single stranded; DNA is double stranded.

 RNA contains ribose sugars where as DNA contains deoxy ribose sugar.
 RNA contains the uracil in place of thymine. Uracil lack methyl group present in
thymine.
 RNA molecules are generally much shorter than DNA molecules.
Three types of RNA

1. Messenger RNA or mRNA

It is a kind of single strand RNA molecule which is complementary to sense strand of DNA
molecule. It is produced by transcription of structural genes in the DNA sequence. It carries the
genetic message from the chromosome to the site of protein synthesis [Link]. mRNA
molecule corresponds to each gene that is expressed.
2. Transfer RNA or tRNA

Transfer RNA is a kind of RNA molecule consists of 75 mucleotides and it becomes smallest
of all RNA molecules. They carry the activated aminoacids to the ribosome for protein
synthesis. There is a specific tRNA for each of the twenty aminoacids. The 5‟ end of tRNA
have poly G and it is phosphorylated. tRNA contains many unusual bases between 7 and 15
per molecule. tRNA is folded into a clover leaf pattern. It has five following special region as
follows.

[Link] end : The base sequence in 3‟ end of all tRNA is CCA. The activated amino acid is
attached to 3‟ hydroxyl group of the terminal adenosine.

ii.TΨC arm: Involved in the binding of the tRNA to ribosome.

iii. Anticodon loop : It consists of seven bases and codon Recognition site. Codon recognition
site is complementary to codons of mRNA.

[Link] arm : It is the site for the recognition of amino acid activated enzymes

v. Extra arm : Some tRNA have extra arm.

FUNTIONS of tRNA

i. tRNA picks up a specific activated amino acid (charging) from the aminoacid pool in the
cytoplasm.
ii. The aminoacid is then transferred to the ribosome where proteins are synthesized.
iii. The attachment with ribosome depends upon the codons in the mRNA and anticodons in
the tRNA.
iv. Finally it transmits its amino acid to the new polypeptide chain.
3. Ribosomal RNA or rRNA : Ribosomal RNA is a ribonucleic acid present in the ribosomes
and hence it is called ribosomal RNA. It is a kind of RNA molecule serving as a major
component of ribosomes. It does not contain a genetic message.E .Coli has three kinds of rRNA
i.e., 23s, 16s, 5s. rRNA is transcript of rRNA genes.

As it becomes evident that the genes controlled the structure of polypeptides, attention
focused on how the sequence of four base pairs in DNA control the sequence of 20 amino acids
found in proteins.

GENETIC CODE

In the DNA and RNA there are four types of nucleotide or bases viz., A, G, T, C and A,
G, U C respectively. If it is assumed that each base codes (singlet) for one amino acid, then
only four amino acids can be coded. If two bases together (doublet) are responsible for one
amino acid, then they will code for 42 = 16 amino acids. If three bases together code (triplet)
for an amino acid then 43 = 64 amino acids could be coded. As the essential amino acids in a
biological system are 20 in number, the possibility of one or two bases coding for each amino
acid is remote.
Crick and Brunner (1961) suggested that the genetic code might be a triplet code,
involving three nucleotide bases to code for an amino acid. Nirenberg and Mathaei (1961)
experimentally proved that a single amino acid is determined by sequence of three nitrogen
bases, known as triplet code.
Definition of genetic code

The genetic code is the relationship between the sequences of bases in DNA (on its
mRNA) and the sequence of amino acid in the protein. The sequence of three nucleotides in
mRNA that specifies a particular amino acid in the protein synthesis is called genetic
[Link] are 64 codons, of these 61 are sense codons i.e. each coding a specific amino acid
and the remaining 3 are non-sense codons ( code for termination of the message)

Anticodon : The anticodon is a sequence of three base pairs of the tRNA, complementary to
the matching three base pairs of codon on the mRNA.
It is also matched to a specific amino acid in a tRNA molecule. The anticodons are attached
to the codons.

Pattern of genetic code

1. Codons for the aromatic amino acids begin with uracil

• UUC & UUC -Phenyl alanine (Phe)
• UGG -Tryptophan (Trp)
• UCU & UCC - Serine (Ser)
2. Codons for amino acids that form amides begin with Guanine and Adenine
• GAU & GAC -Asparagine (Asp)
• GAA & GAG -Glutamine (Glu)
3. Many of the synonymous codon specifying the same amino acid
• The first two bases of the triplet code are constant, while the third varies, being less
specific.
• CCU, CCC, CCA, CCG -proline (pro)
• GGU, GGC, GGA, GGG-Glycine (Gly)
Co-linearity between Gene and protein

Benzer revealed that there is a linear correspondence between a gene and its
polypeptide products. This co-linearity gives the clue that specific arrangement of
nitrogenous bases in DNA determines the specific sequence of amino acid in protein. So the
genetic information is written by four-letter language of DNA nitrogen base.

Characteristic features of genetic code

1. Triplet

There are 20 kinds of amino acids in the cytoplasm but only four kinds of nitrogenous
bases. A singlet code is inadequate because it codes for only 4 amino acids and also a doublet
code is also inadequate since it codes for 4x4 =16 amino acids only. A triplet code is only
adequate since it codes for 64 (4x4x4) amino acids. A group of three bases that codes for one
amino acid is called codon. Amino acids coded by the 64 possible codons of the triplet code.

First base Second base position Third base

Position 5’ end Position 3’ end

U C A G
UUU Phe UCU Ser UAU Tyr UGU Cys U
UUC Phe UCC Ser UAC Tyr UGC Cys C
U
UUA Leu UCA Ser UAA Stop UGA Stop A
UUG Leu UCG Ser UAG Stop UGG Trp G
CUU Leu CCU Pro CAU His COU Arg U
CUC Leu CCC Pro CAC His CGC Arg C
C
CUA Leu CCA Pro CAA Glu CGA Arg A
CUG Leu CCG Pro CAG Glu CGG Arg G
AUU Ile ACU Thr AAU Asn AGU Ser U
AUC Ile ACC Thr AAC Asn AGC Ser C
A
AUA Ile ACA Thr AAA Lys AGA Arg A
AUG Met ACG Thr AAG Lys AGG Arg G
G GUU Val GCU Ala GAU Asp GGU Gly U
GUC Val GCC Ala GAC Asp GGC Gly C
GUA Val GCA Ala GAA Glu GGA Gly A
GUG Val GCG Ala GAG Glu GGG Gly G

2. Codons have degeneracy

The occurrence of more than one codon per amino acid called degeneracy. The amino acids
like methionine and tryptophan has only one codon each. Whereas, the amino acids leucine,
serine and arginine have six codons each. The codons that specify the same amino acid are
called synonyms. For example the codons CAU and CAC code for histidine and most of the
synonyms differ only in the last base of the triplet. The degeneracy helps for minimizing the
deleterious effect of mutation.
3. Code is non-ambiguous. It means that a particular codon always code for the same amino
acid. But AUG code for methionine and start codon.

4. Genetic code in non-overlapping

It means that no single base can take part in the formation of more than one codon.

5. Genetic code is commaless

The genetic code is commaless, which means that no codon is reserved for punctuations.

6. Start and stop codon (initiation and termination codon)

UAG (Ochre), UAA (Amber) and UGA (opal) are only the three codons that do not specify
amino acid. They designate chain termination. These codons are not read by tRNA molecules
but by specific proteins called release factors. The codon AUG (methioine) and GUG (valine)
act as start codon for translation.
7. Genetic code is universal

With one or two exceptions the genetic code is same or nearly same in all the organisms. The
codon UGA codes for tryptophan in human mitochondrial system. Whereas, UGA is a
termination codon for non-mitochondrial system. Mutation that produces chain – termination
triplets within genes is called as non-sense mutation whereas mis-sense mutation cause change a
triplet to another triplet specifying a different amino acids.

8. Codons have wobbling

A codon of mRNA is recognized by the anticodon of a tRNA base of codon forms a Watson –
Crick type of base pair with a complementary base on the anticodon. The codon and anticodon
are antiparallel in base pairing. Some tRNA molecules can recognize more than one codon. For
example, the yeast alanine tRNA binds to three codons: GCU, GCC and GCA. This is explained
by Wooble hypothesis. Crick (1966) proposed wobble hypothesis (Wobble means move
unsteadily or non-specific) to explain how one tRNA recognise two codons.
Hypothesis states that only first two bases of a codon have a precise pairing with the bases of
the anticodon (base pairing rules), while the third one may wobble (non-specific).

Significance of Wobble hypothesis

• Wobbling allows economy of the number of tRNA molecules.

• Wobbling enable to minimise mutational lethality.
 • Hydrogen bonding between bases in anticodons and codons more stringent for first two
bases of the codon and less stringent for third base.
• The post transcriptional enzymatic modification of several tRNAs contain inosine base
which can pair with A, U or C.
5’ base in anticodon 3’ base in codon in the mRNA

G U or C
C G
A U
U A or G
I (inosine) A, U, C

The central dogma

The sequence of amino acids in a protein is determined by the base sequence of DNA, through
the m RNA. The flow of genetic information is

DNA ----- Transcription ---------mRNA ---------Translation -------- Protein

mRNA -------Reverse Transcription -------------- DNA

I. Transcription

Transcription is the first step in gene expression. Transcription is the process of creating a
complementary RNA of a sequence of DNA. Transcription is the synthesis of mRNA from a
DNA template. It is like DNA replication in which a DNA strand is used to synthesize a strand
of mRNA. Only one strand of DNA is copied. After transcription, the DNA strands [Link]
stretch of DNA transcribed into an RNA molecule is called a mRNA transcript.

Steps involved in transcription

The synthesis of mRNA (transcription) takes place in three stages (i) initiation (ii) elongation
and (iii) termination.

1. Initiation of transcription

First step in transcription is unwinding of DNA segment. Of the two strands, one strand
act as sense strand which is used as template for mRNA synthesis. The other strand viz., anti-
sense strand remain untranscriped. RNA synthesis does not require any primer to start. The
transcription starts at specific sites, called promoters on the DNA template. The promoter sites
consist of about 40 base pairs (about 140 Å). RNA synthesis takes place in 5‟ to 3‟ direction
like DNA synthesis.

The DNA strand is read from 3‟ to 5‟ direction by RNA polymerase. RNA polymerase
is a holoenzyme, consists of two major parts viz., the core enzyme and sigma factor. The core
enzyme and sigma factor join to from holoenzyme and initiate the transcription process.
The sigma factor activates RNA polymerase to recognize promoter sequences. RNA polymerase
binds to specific base sequence in the DNA called a promoter. The promoter identifies the start
of a gene, which strand is to be copied, and the direction that it is to be copied. The sigma
subunit dissociates from the holoenzyme after the new RNA chain is started. The core
polymerase continues to transcribe the DNA template.
RNA chains start with pppA or pppG. That is, the new RNA chain has a triphosphate
group at its 5‟ terminus and a free hydroxyl group at its 3‟ terminus. In contrast with DNA
synthesis, primer is not needed and RNA chains can be formed de novo.

ii. Elongation of transcription

Core enzyme catalyzes the mRNA chain elongation. The RNA polymerase moves along the
DNA template strand in the 3‟to 5‟ direction because the template strand is antiparallel to the
newly synthesized RNA strand. The same RNA polymerase molecule synthesize RNA strand.
The transcribed region of DNA remains its double – helical conformation as the next section of
DNA unwinds. The maximum rate of elongation is about 50 nucleotides per second.
In contrast to DNA polymerase, RNA polymerase does not edit the nascent polynucleotide
chain. Hence, the fidelity of transcription is much lower than that of replication. The error rate
of RNA synthesis is of the order of one mistake per 104 or 105. The lower fidelity of RNA
synthesis can be tolerated because a cell synthesizes many RNA transcripts of a gene.

iii. Termination of transcription

The DNA template contains stop signals for transcription. Before the termination site, GC rich
region is followed by an AT rich sequence. The most distinctive feature of termination
sequences is the two fold symmetry of their GC rich region. Hence, the RNA transcript of this
region is self-complementary and so, it can base-pair to form a hairpin structure. In addition, the
nascent RNA chain ends with several U residues, which are specified by a series of A bases in
the AT-rich region of the DNA template. These structural features cause RNA polymerase to
pause when it encounters such a signal. The rho protein participates in the termination of
[Link] binds to RNA polymerase to stop the [Link] mRNA produced is
called a mRNA transcript.

Processing the mRNA Transcript

In eukaryotic cells, the newly-formed mRNA transcript (also called heterogenous nuclear RNA
or hnRNA) must be further processed before it can be used. A cap is added to the 5‟ end and a
poly-A tail (150 to 200 Adenines) is added to the 3‟ end of the molecule. The newly-formed
mRNA has regions of exons and introns. Introns do not contain a genetic message. These
introns must be removed. The remaining portions of mRNA after removal of introns are called
exons. They spliced together to form a mature mRNA transcript

II. Translation

Translation is the process where ribosomes synthesize proteins using the mature mRNA
transcript produced during transcription. Translation is directed by the mRNA and
accomplished with the help of a ribosome, rRNAs and tRNA. Transfer RNA, translates the
sequence of codons on the mRNA strand. tRNAs continue to add amino acids to the polypeptide
chain until they reach a stop codon on the mRNA. The ribosome then releases the completed
protein into the cell.
Ribosomes – the site of protein synthesis

The ribosome of eukaryotic cells is larger. It has sedimentation coefficient of 80 S. It dissociates

into 60 S and 40 S sub units. The ribosomes of [Link] has a sedimentation coefficient of 70 S. It
dissociated into a large subunit (50S) and a small subunit (30 S). Many ribosomes can
simultaneously translate an mRNA. This increases the efficiency of utilization of the mRNA.
The group of ribosomes bound to an mRNA is called a polyribosome or a polysome. Each
ribosome operate independently, synthesizing a complete polypeptide chain. The maximum
density of ribosomes on mRNA is about one ribosome per eighty nucleotides. Ribosomes near
the 5‟ end of the mRNA have shortest polypeptide chains.

Steps in Translation

1. Initiation of translation

The amino acids are activated by ATP. In the first step, the carboxyl group of the amino
react with ATP, forming amino acyl adenylate and releasing pyrophosphate. The amino acyl
adenylate reacts with tRNA and forms amino acyl- tRNA. The attachment of an amino acid to a
tRNA is important because amino acids themselves cannot recognize the codons on mRNA.
Amino acids are carried to the ribosomes by specific tRNAs which recognize codons on
mRNA. (iii)Thus, the specific tRNAs acts as adaptor molecules.

Protein synthesis is initiated by formyl methoinine tRNA. N-formyl methionine (f Met)

is a modified methionine which has a formyl group attached to its terminal amino group. Protein
synthesis starts with the association of mRNA, a 40 S ribosomal subunit, and formyl methionyl
– tRNA to form a 40 S initiation complex. The formation of this complex requires GTP and
three protein factors viz., 1F-1, 1F-2, 1F-3. IF-3 mediates the binding of mRNA to 40 S
subunits and 1F1, and 1F-2 enhance the binding of initiator tRNA to the mRNA – 40S subunit
complex.

A 60 S ribosomal subunit then joins a 40 S initiation complex to form an 80S initiation

complex. The 80S initiation complex is ready for the elongation phase of protein synthesis.
Ribosome contains two sites A and P. The f Met – t RNA molecule occupies the P (peptidyl)
site on the ribosome. The other site for a tRNA molecule on the ribosome, the A (aminoacyl)
site is empty. The anticodon of f Met – tRNA pairs with the initiating AUG (or GUA) codon on
mRNA. Thus, the reading frame is defined by specific interactions of the ribosome and of Met
RNA within RNA.

First the mRNA attaches to the small subunit ribosome. Six bases of the mRNA are
exposed. A complementary tRNA molecule with its attached amino acid (methionine) base
pairs via its anticodon i.e. UAC with the AUG on the mRNA in the first position P. Another
tRNA base pairs with the other three mRNA bases in the ribosome at position A. The first
tRNA (without its amino acid) leaves the ribosome.

2. Elongation of Translation

Elongation depends on elongation factors viz., eEF-1 and eEF-2. The mRNA is
positioned in such a way that next codon is translated. The second tRNA molecule moves into
position P. Another tRNA molecule pairs with the mRNA in position A bringing its amino acid.
The enzyme peptidyl transferase forms a peptide bond between the two amino acids. The
ribosome moves along mRNA in 5‟ to 3‟ direction. This brings the next codon to A site and
next tRNA attached with its anticodon. In this way, polypeptide chain elongates and new
polypeptide is formed until a stop codon is reached.
3. Termination of translation

Termination occurs when any one of the stop codon moves into A site. Termination
codon not recognised by tRNAs. Termination of polypeptide is depending on release factor
eRF.
Three stop codons are recognized by soluble proteins called release factors (RFs)
eRF 1 – UAA and UAG
eRF 2 – UAA and UGA
The presence of a release factor in the A site stop the activity of peptidyl transferase. The RF
releases the protein from ribosome and signals the ribosome to leave the mRNA. The 80 s
ribosome then dissociates into 40 S and 60S subunits as the prelude to the synthesis of another
protein molecule.
Lecture 33- Regulation of gene expression – Operon model of Jacob and Monad;
Structural genes and regulator genes. Cistron, muton and recon

Gene regulation: The study of on-off mechanism of protein synthesis; also known as
regulation of gene action, regulation of gene expression and regulation of protein synthesis.
Positive regulation: Enhancement of transcription by an effector molecule through activation
of promoter. Eg. Lac operon of [Link].
Negative regulation: Inhibition of transcription by repressor through inactivation of promoter.
Eg. Trp operon
Repressor: A protein molecule which prevents transcription. The process of inhibition of
transcription is called repression. Eg. Trp operon, tryptophan acts as repressor.
Inducer: The substance which allows the initiation of transcription eg. Lactose in lac operon.
The process of initiation of transcription is known as induction.
Co-repressor: A combination of repressor and metabolite which prevents protein synthesis.
Such process is termed as co-repression.
Inducible enzyme: An enzyme whose production is enhanced by adding the substrate in the
culture medium.
Repressible enzyme: An enzyme whose production is inhibited by adding an end product.
Constitutive enzyme: An enzyme whose production is constant irrespective of metabolic state
of the cell.
Effector: The molecule which acts as an inducer or co-repressor in the operon model of [Link].
Operon model: A group of closely linked genes which acts together and code for the various
enzymes of a particular biochemical pathway.
Structural genes: The genes in lac operon, which control the synthesis of protein through
mRNA.
Promoter gene: A gene in lac operon of [Link] which initiated transcription. It is located
between regulator and operator genes.
Operator gene: A gene found in lac operon and directs the synthesis of repressor protein
molecule.
Regulator gene: A gene operon model of [Link] which controls the function of structural genes,
 Each cell of the living organisms contains thousands of genes. But all genes do not
function at a time. Genes control the phenotypic expression of characters through the production
of specific enzymes. The synthesis of particular enzyme is depending upon the requirement of
the cell. Thus, there exists an on-off system which regulates protein synthesis in all living cells.
The study of this on-off mechanism is called regulation of gene expression.

The gene expression is regulated at many different levels. They are

 Transcriptional level
 mRNA processing
 mRNA turn over
 translation level
 enzyme function
Most of the data indicate that regulation of transcription is the most important mode of control
of gene expression.

Synthesis of enzyme depends mainly on two factors. In catabolic pathway, the synthesis of
enzyme depends on the availability of the molecule to be degraded. In biosynthetic pathway
the synthesis of an enzyme governed by end product.

MECHANISM OF GENE REGULATION

1. Negative regulation

– In negative control, the regulator protein acts as the repressor and prevents
transcription of structural genes by binding with the operator.
– The main controlling factor is the operator gene.
– Gene expression is possible only when the operator is free.
– Absence of end product increases synthesis and presence of product decreases
synthesis of repressor protein.
Eg. Presence of glucose inhibits the production of enzyme β-galactosidase.

2. Positive regulation

• The regulator protein functions as an activator and increases the enzyme synthesis.
• The activator binds to initiator site on DNA and enhances the transcription of
structural genes
• Presence of a product will enhance the synthesis of enzyme.
Eg. Presence of lactose increases the production of enzyme β-galactosidase.

3. Auto Regulation

Protein synthesized by a gene directly inhibits the transcription by the same gene when the
protein is produced in high concentration

4. Coordinated Regulation

 When several enzymes act in a sequence in a single metabolic path way, the regulation
is brought by the inhibition of synthesis of either all or none of the enzymes.
 Inhibition of all enzymes results from the control of synthesis of single polycistronic
mRNA molecule encoding all the enzymes.
[Link] operon

Traditionally, the gene has been defined as the unit of genetic material controlling the
inheritance of one phenotypic characteristic or one trait and also believed that gene was not to
be sub divisible by mutation or recombination. Today the gene is precisely defined as the unit
of genetic material coding for one polypeptide.

Types of gene expression

1. Constitutive expression

Some genes are essential and necessary for life and therefore are continuously expressed.
These genes are called housekeeping genes.
2. Induction and repression

The expression levels of some genes fluctuate in response to the substrate.

The synthesis of some enzymes can be increased many times by the presence of substrate are
inducible enzymes. The genetic system for the synthesis of such enzymes is inducible system.
On the other hand, presence of substrate reduces the synthesis of an enzyme. Such enzymes
are repressible enzymes and the system is repressible system.

Differences between prokaryotes and eukaryotes gene regulation

i. Prokaryote gene expression typically is regulated by an operon, the collection of
controlling sites adjacent to protein-coding sequences. Each eukaryotic gene has its
promoter and enhancer elements, but operons are not known to occur.
 ii. Eukaryotic gene regulation is more complex because transcription and translation are
not coupled. Simultaneous transcription and translation in prokaryotes.
iii. Eukaryotic genes are switched off state for transcription and prokaryotic genes are
switched on state for transcription.
iv. There is no processing of mRNA in prokaryotes unlike eukaryotes.
v. The mRNA produced is polycistronic in nature, whereas it is monocistronic in
eukaryotes.
vi. Post transcriptional and post translational regulation occur in eukaryotes.
vii. In prokaryotes, repressor protein act as regulator of gene expression and in
eukaryotes, activator RNA acts as a regulator.
Operon
An operon refers to a group of structural genes whose transcription is regulated by the
coordinated action of regulator gene and an operator [Link] usually controls an
important biochemical process. They are only found in prokaryotes.
Operon structure

The operon model

[Link] and [Link] (1961) proposed the operon model to explain the regulation of genes
coding the enzymes required for lactose utilization in [Link]. The operon is a co-ordinated unit
of the gene expression. The operon consists of structural genes, the operator, regulator and
promoter.

Lac operon (Enzyme induction)

Structural genes

The lac operon of [Link] consists of three structural genes namely z,y and a . These structural
genes transcribe a single polycistronic mRNA molecule. This mRNA molecule controls the
 synthesis of three different enzymes viz., β-galactosidase, galactosidase permease and
galactosidase transacetylease. All the above three enzymes are involved in breakdown of
lactose. The function of all the structural genes is regulated by two controlling elements
namely regulator and operator. Thus, the main function of structural genes is to control
synthesis of protein through mRNA.

Operator gene

The operator is usually located between the promoter and the structural genes. In lac operon of
[Link], operator is located contiguous to the structural genes. It is the binding site for the
protein called repressor. When the repressor is bound to the operator, transcription of the
structural genes cannot occur, because the binding of the repressor to the operator strictly
prevents RNA polymerase from binding at the promoter site.

Promoter gene

The promoter gene is always located contiguous with or even overlapping with operator
sequence or operator. The promoter segment is a place where mRNA polymerase enzyme
binds with DNA. The main function of promoter gene is to initiate mRNA transcription. The
promoter starts mRNA transcription only when operator is free or when repressor is not bound
to the operator. The binding of repressor with operator inactivates the promoter gene and
prevents transcription.

Regulator gene

The regulator gene is located either on one end of the operon or away from the operon. The
function of the regulator gene is to synthesis a protein called repressor. The repressor may be
either active or inactive.

In the case of an inducible operon, the free repressor binds to the operator and turning off the
transcription. In the case of reversible operon, the repressor is inactive and cannot bind to the
operon there by transcription of structural genes in the operon is turned on.

Mechanism of gene regulation in lac operon

The lac operon consists of three structural genes (lac Z, lac Y and lac A), each one involved in
processing the sugar lactose
1. lac Z- gene codes for β-galactosidase - hydrolyses lactose into glucose and galactose
 2. lac Y - Lactose permease - permits entry of lactose from the medium into the bacterial cell.
3. lac A - Thiogalactoside transacetylase - transfers an acetyl group from acetyl co-enzyme A
to β - galactosidase.

Near the operon on the DNA is a regulatory gene called the “I” gene. This codes for the
repressor protein.
Lac operon turn off: In the absence of lactose in the medium, the regulator gene produces the
active repressor molecule. These repressor molecules will bind to the operator and it strictly
prevents the RNA polymerase from binding to the adjoining promoter. Thereby synthesis of
enzymes by structural genes involved in lactose metabolism viz.,galactosidase, galactosidase
permease and galactosidase transcetylase were switched off.
Lac operon turn on: When the lactose (effectors molecule) is added to the medium, which act
as inducer, will bind to the repressor and become repressor-inducer complex. This complex is
inactive in nature, which cannot bind to the operator. The operator is now free from repressor
and RNA polymerase will bind with promoter region and start the transcription of structural
genes involved in lactose metabolism.
Control of the lac operon

Four situations are possible

1. When glucose is present and lactose is absent, the E. coli does not produce
β-galactosidase

A repressor protein is continuously synthesised. It sits on a sequence of DNA just in front

of the lac operon, the Operator site. The repressor protein blocks the Promoter site where the
RNA polymerase settles and stops transcription.

2. When both glucose and lactose are present, E. coli does not produce
β-galactosidase

A small amount of a sugar allolactose is formed within the bacterial cell. This fits onto the
repressor protein at another active site (allosteric site). This causes the repressor protein to
change its shape (a conformational change). It can no longer sit on the operator site. RNA
polymerase can now reach its promoter site.

3. When both glucose and lactose are absent, the E. coli does not produce β-
galactosidase

A repressor protein is continuously synthesised. It sits on a sequence of DNA just in front

of the lac operon, the Operator site. The repressor protein blocks the Promoter site where
the RNA polymerase settles and stops transcription.
 4. When glucose is absent and lactose is present, the E. coli does produce β-
galactosidase

Another protein is needed, an activator protein. This stabilises RNA polymerase. The
activator protein only works when glucose is absent. In this way E. coli only makes
enzymes to metabolise other sugars in the absence of glucose.

Trp operon (Enzyme repression)

[Link] is capable of producing the enzymes necessary for the biosynthesis of amino acids as
well as other macromolecules. If tryptophan is present in sufficient quantity in growth medium,
the enzymes necessary for its synthesis are not produced. A series of five enzymes encoded by
five contiguous genes are involved (trp E, D, C, B and A). In the presence of tryptophan, all
the enzymes are coordinately repressed and none of the enzymes are produced.
Modern concept of gene

Seymour Benzer (1955) described the fine structure of gene and introduced three terms-
Cistron, Recon and Muton. Now the gene can be defined as the unit of genetic material coding
for one polypeptide. So the gene can also be called as cistern.

Cistron (unit of genetic function)

Cistron is the segment of DNA specifying a single polypeptide chain. It has information for
synthesis of specific protein. It is responsible for expression of character or trait. It may be
several base pairs (bp) long. It is functional unit of hereditary element.

Recon (unit of recombination:

The segment of DNA which is capable of undergoing crossing over and recombination during
meiosis is called recon. It is few to many base pairs long. There are many sites in a cistron that
undergo recombination.

Muton (unit of mutation):

The smallest segment of DNA which undergoes mutation is called muton. It is the unit of
DNA which when changed or mutated produces a different phenotypic trait. There are many
sites within cistron where mutations can occur. 4. Unit of mutation is smaller than a cistron. It
consists of one to few nucleotides.

The cistron contains so many mutons and recons, the smallest unit of mutation or
recombination.

Transposable elements (TE) / Mobile Elements

Transposition is the movement of genetic information from one chromosomal location, the
donor site, to another location, the target site. DNA sequences that can change their genomic
location intragenomically either autonomously or non-autonomously are called as transposable
elements.

 Autonomous transposable elements have the ability to excise and transpose

themselves & corresponding non-autonomous elements.
 Non-autonomous transposable elements do not have the ability to transpose. They
transpose only when an autonomous member of the same family present in the genome.
 First discovered by Barbara McClintock in 1940's, It is also called as jumping genes.
Her work was validated when these elements were later discovered in bacteria in the 1960's.

General Features of Transposable elements

i. Ubiquitous in eukaryotes and prokaryotes.

ii. Transposable elements can move from chromosome to chromosome or plasmid to
chromosome, or chromosome to plasmid.
iii. In both eukaryotes and prokaryotes, transposable elements insert into new locations with
which they have no sequence homology (i.e. nonhomologous recombination).
iv. Cause mutations by insertional inactivation, either into coding region or into regulatory
sequences, altering expression levels.
v. Transposable elements also cause chromosome rearrangements. This has had enormous
impact in evolution of genomes.

Mechanisms for transposition

1. Conservative transposition: The mobile element itself moves from the donor site into
the target site
2. Replicative transposition: The mobile element moves a copy of itself to a new site via
a DNA intermediate
3. Retrotransposition: The mobile element makes an RNA copy of itself which is
reversed-transcribed into a DNA copy which is then inserted (cDNA)

Transposable Elements in Prokaryotes

1. Insertion Sequences (IS) : IS elements contain only genes required to mobilize the
element and insert the element at a new location. Range in size from 768 bp to 2.5 kb
2. Transposons: Similar to IS elements but are more complex structurally and carry additional
genes that are not required for [Link] in size from 2.5 kb to 7.0 kb

Examples of Transposable Elements in Eukaryotes

1. Ac – Ds System in Maize: Barbara McClintock has been awarded Nobel Prize in

Physiology or Medicine in 1983 for the discovery of jumping genes in maize. She stated
that if Ac elements are absent the transposition does not occur, the genes express their
normal phenotype (C’ gives colourless kernels and C lets the colour to express). On the
 other hand, if transposition occurs in some cells, the chromosome undergoes breakage and
the integration of the Ds element in the gene, makes it mutant resulting in the cell and all
its daughter cells to show pigmentation. If the mutation occurs early in development, more
pigmented (variegated) cells would be produced and vice – versa.

2. Transposable Elements in Humans: The major families of transposable elements

constituting the human genome include LINEs (size 1000 to 6000 base pair long with
around 850000 copies constituting a total of 21% of the genome) and SINEs (size 100 to
500 base pair long with 1500000 copies thereby forming 13% of the total genome size).
These together constitute nearly 44% of the genome. Insertion of LINEs and SINEs in
the genomes has been reported to cause numerous disorders like haemophilia, Duchene
muscular dystrophy, cancers, neurofibromatosis etc.
Lec 34 : Mutation – characteristics of mutation – micro and macro mutation – ClB
technique - molecular basis of mutation- Transition and transversion; major physical and
chemical mutagens

Mutation : Mutation is the sudden heritable change in the phenotype of an organism. In

molecular term, mutation is the permanent change in the number or sequence of nucleotides.
Spontaneous mutation: Mutations that occur in nature.
Induced mutation: Mutations that are induced by the treatment of mutagenic agents.
Macro mutation: Mutation with distinct morphological changes in the phenotype. Usually
observed in qualitative characters.
Somatic mutation: A mutation in somatic tissues or cells.
Forward mutation: Any change from the wild type allele to mutant allele.
Reverse mutation: A change from mutant allele to wild type.
Hot spots: Highly mutable sites within a gene.
Mutant: The product of mutation. It may be a genotype, a cell or a polypeptide.
Mutagen: Physical or chemical agents which greatly enhance the frequency of mutation.
Transition: Substitution or replacement of one purine by another purine (replacement of
adenine by guanine and vice versa) or a pyrimidine by another pyrimidine (replacement of
thymine by cytosine and vice versa).
Transversion: Substitution or replacement of a purine (adenine or guanine) by a pyrimidine
( Thymine or cytosine) or vice versa.
Tautomerization: When the hydrogen atoms in a DNA molecule gets shifted from one position
to another in a purine or in a pyrimidine, the process is called as tautomerization and the new
product is called as tautomer.
Frameshift mutation: Mutations which arise due to addition or deletion of nucleotides in
mRNA.
Nonsense codons: Mutations which have nonsense codons. Nonsense codons do not code for
any amino acids.
Missense codons: Mutations which have missense codons. Missense codons code for different
amino acids.
Point mutation: Gene mutation at molecular level.
Mutagenesis: The mechanisms by which spontaneous or induced mutations occur.
LD 50: A dose of mutagen that kills 50% of the treated individuals.
Mutation

Mutation is a sudden heritable change in a specific character due to change in the DNA
sequence of a gene. In molecular term, mutation is the permanent change in the number or
sequence of nucleotides which results due to change in DNA bases in nuclear or cytoplasmic
DNA or due to chromosomal aberrations.

It was first discovered by Wright (1791) in male lamb with short legs. It was reported by
Hugo de Vries (1900) in Oenothera lamarckiana (evening primrose). The term mutation was
introduced by Hugo de Vries Hugo de vries (1901) from the latin word mutare – “to
change”.Mutagenic action of X –rays was discovered by Muller in 1927 on Drosophilla and of
gamma rays and X-rays in 1928 by Stadler in barley.

General characteristics of mutation

i. Mutations are generally recessive and rarely dominant mutations also occur.
ii. Mutations are generally harmful to the organism. A small proportion of them (0.1%)
are beneficial.
iii. Mutations occur at random (i.e) they may occur in any gene or chromosome. But
some genes show higher mutation rates than others. Highly mutable sites are called
mutational hot spots.

iv. Mutations are recurrent (i.e) same gene may undergo mutations repeatedly.
v. Induced mutations often show pleiotropy due to close linkage of mutated gene with
other genes.
vi. Macro mutations which occur in oligogenes are easily identifiable and micro
mutations which occur in poly genes are not easily identifiable.

Classification of mutations

1. Depending upon the size of genetic material involved in the mutation

i. Point mutation : If the mutation affects a single gene causing a change in the allele status
fromdominant to recessive or vice versa, the mutation is called as point mutation. It is also
called as intra genic mutation. Change in allelic status may be from wild type to a new type
called as forward mutation or may be from new type to wild type called as reverse mutation.
ii. Chromosomal mutation : If the mutation induces structural aberration in the chromosomes
such as deletion, duplication, inversion and translocation or numerical aberrations, the mutation
is called as chromosomal mutation.

2. Depending upon the magnitude of phenotypic effect produced

i. Macro mutation : If the mutation result is easily identifiable distinct morphological
changes in the phenotype, the mutation is called as macro mutation. It is called as oligogenic
mutation, as it is observed generally in qualitative characters which are controlled by
oligogenes. Since, the phenotypic change is easily recognizable on individual plant basis;
selection in M2 generation is easy.

ii. Micro mutation: If the mutation result is not easily identifiable and not clearly
distinguishable morphological changes in the phenotype, the mutation is called as micro
mutation. It is called as polygenic mutation as it is observed in quantitative characters which
are controlled by polygenes. Micro mutations are of most economic value in plant breeding
than macro mutation as most of the economic characters are governed by polygenes. In micro
mutation, since the small phenotypic change is not recognized on individual plant basis and
detected only in a group of plants, selection is carried out on M3 or later generations only.

3. Depending upon the survivability level of mutated individuals

Lethal: If the mutation kills all the individuals carrying the mutated gene, the mutation is
called as lethal mutation. Lethal mutation reduces the viability of the individuals completely
and hence all the individuals are killed. Dominant lethals (AA) do not survive in
homozygous or heterozygous condition and hence cannot be studied. Recessive lethal (aa)
do not survive in homozygous condition alone. E.g Allina chlorophyll mutation.
Sub-lethal: If the mutation kills more than 50% of the individuals carrying the mutated
gene, the mutation is called as sub-lethal. Sub lethal mutation reduces the viability of the
individuals to a greater extant but not completely. Hence, most of the individuals are killed.
Sub-vital: If the mutation kills less than 50% of the individuals carrying the mutated gene,
the mutation is called as sub- vital mutation. Sub-vital mutation reduces the viability of the
individuals to a limited extent and hence lesser no of individuals are killed.

Vital: If the mutation does not kill any of the individuals carrying the mutated gene,
mutation is called as vital mutation. Vital mutations do not reduce the viability of the
individuals and hence all the individuals survive. Vital mutations occur in a much lower
frequency (0.1% of all the mutations) as compared to other three types.

Vast majority of mutations are lethal, sub-lethals, and sub-vital and are of little value in
crop improvement and sometimes have considerable academic interest. Vital mutations are
of high value in crop improvement.

4. Based on types of cells in which mutation occurs

(i) Germinal mutation

If the mutation occurs in the generative cells (reproductive cells) of an organism, the
mutation is called as gametic mutation and the mutation is passed on to the next generation,
via the gametes.
(ii) Somatic mutation
If the mutation occurs in the somatic cells of an organism, the mutation is called as somatic
mutation. It is of two types.
(a) Non heritable somatic mutation
In sexually reproducing organisms, if the somatic mutation occurs, the mutation lasts up to
the end of the generation and is not inherited to the next generation.
(b) Heritable somatic mutation (Bud mutation)
In asexually reproducing organisms, if the somatic mutation occurs, the propagating part of
the somatic cells of the mutation is called as heritable somatic mutation and the mutation is
inherited to the subsequent generation through vegetative propagation. Such somatic
mutations occurring in vegetative propagation is called as bud mutations. Since, in general
buds are the core vegetative propagating part.

5. Based on the source of origin or presence or absence of artificial causal factors

(i) Spontaneous mutation

If the mutation occurs naturally without the artificial treatment by physical or

chemical agents, the mutation is called as spontaneous mutation. Frequency of spontaneous
mutation is generally very with range of one in 10 lakhs. i.e 10-6. However, some loci
undergo spontaneous mutations at high frequency. E.g R locus in maize with a frequency of
4.92 x 10-4, while Wx locus is highly stable. Spontaneous mutation of certain genes is
influenced by the genetic background in which it is present.
 Certain intrinsic and extrinsic factors produce spontaneous mutation. Genetic
instability due to hybridity or polyploidy or due to specific genes promoting chromosomal
sickness in mitosis or meiosis and physiological conditions are the intrinsic factors involved
on spontaneous mutation. Extrinsic (external) factors causing spontaneous mutation are
nutrition, temperature, naturally occurring radiations are chemicals and very high oxygen
pressure.

ii) Induced mutation

If the mutation occurs due to the artificial treatment by physical or chemical

mutagenic agents, the mutation is called as induced mutation. Induced mutation produces
new allele similar to spontaneous mutations. But in contrast to spontaneous mutation,very
high frequency of mutation can be induced through the mutagenic agents at desired level.
Hence induced mutations are more useful in crop improvement. Muller produced mutants in
Drosophila using X-rays

6. Based on type of chromosomes in which it occurs

i. Autosomal : mutation occur in autosomes

ii. Sex linked mutations : mutations occur in sex chromosomes

7. Depending upon the location of the genetic material of the cell involved in mutation.
(i) Nuclear mutation
If the mutation occurs in the nuclear genetic material, the mutation is called as
nuclear mutation.
(ii) Cytoplasm mutation
If the mutation occurs in extra nuclear genetic material, the mutation is called as
cytoplasmic or plasma gene mutation. Cytoplasmic genetic material is present in mitochondria
and chloroplast, which also controls the expression of certain characters of the organism like
cytoplasmic male sterility.
[Link] on the occurrence

i. Recurrent mutation : Occurrence of same type of mutant repeatedly in different individuals

of same population.

ii. Non-recurrent: Very rare occurrence

Molecular basis of mutations

In any organism, triplet codon consisting of three nitrogenous bases in the DNA codes
for an amino acid and a specific combination of triplet codons produce several amino acids
which unite through peptide linkage to produce a protein. Proteins (which include enzymes)
decide the phenotype of the organism. Hence, any change in triplet codon (a single or no of
bases) causes a change in the sequence of amino acids in a protein which results in altered
phenotype. A heritable change in Nitrogen base is the ultimate cause of mutation. Chromosomal
aberrations (change in no. or segment) occurring due to mutation also results in the change in
the sequence of amino acids or in the absence of entire protein.

Depending upon types of nitrogenous base level changes molecular basis of mutation can be
classified as

I. Point mutations without frameshift

1. Base substitution: When one nitrogenous base in a DNA molecule is replaced by another
one, it is called as base substitution. It is of two types.

(a) Transition: When a purine is replaced by another purine (replacement of adenine by

guanine and vice versa) or a pyrimidine by another pyrimidine (replacement of thymine by
cytosine and vice versa) the base substitution is called as transition.

Four changes are possible in A G

and T C

(b) Transversion : When a purine (adenine or guanine) is replaced by a pyrimidine ( Thymine

or cytozine or vice versa), the base substitution is called as transversion.

A T or C
G T or G

II. Point mutation with frame shift

If the no. of bases added or lost is not a multiple of three, the sequences of all the
triplet codons beyond the point of insertion or deletion are altered and all the codons code
for a different amino acid. Thus the reading frame of the subsequent codons is shifted in
such mutations. This type of mutation is called as frame shift mutation. A frame shift
 mutation, changes all the amino acids of the concerned protein, located subsequent to the
addition or deletion of bases. In such cases, the concerned protein becomes non
functioned. Hence, such mutations are much more deleterious than those produced by base
substitution except non sense mutations.

1. Base addition: When one or more nitrogenous bases in a DNA molecule are lost it is called
as base addition.
2. Base deletion: When one or more nitrogenous bases in a DNA molecule is lost, it is
called as base deletion.

Normal sequence of nucleotide

G A C T A T C G A A C A T C A C G A

1 2 3 4 5 6

(Amino acids are symbolised 1,2,3)

Insertion of single nucleotide

(C) resulting in G A C T A T C G A (C) A C A T C A C G
nucleotide 1 2 3 (10) (8) (7)
mutant sequence

Deletion of single
nucleotide (T ) GAC ATC GAA CAT
1 8 9 11

Codons which resulted from frame shift mutations fall into three categories.
1. Sense codons : Which are read or translated the same way as it was before frame shift
mutation.
2. Mis-sense codons : Which code for a different amino acid.
3. Nonsense codons: A base change in codon within the coding sequence that converts
into a stop codon.
3. Transposition
A base pair is lifted from one site and placed in another site of the same reading frame.
It also shift the reading frame from the location of removal through insertion location and
upto termination codon.
4. Tautomerization

Tautomeric shift involves shift in hydrogen atoms in a DNA molecule gets from one
position to another in a purine or in a pyrimidine that alters it from one isomer to another
isomer. The process is called as tautomerization and the new product is called as tautomer.

Normal Adenine : Links with Thymine

Guanine : Links with Cytosine
But due to tautomerism an unusual base pairing.
such as A - C
G–T
may result unusual base paring always cause changes in the character.

Mutagens: The agents that induce mutation are known as mutagens. It may be physical or
chemical.
I. Physical mutagens
Physical mutagens are radiations. Radiation causing mutations are of two types
(1) Ionizing radiation (2) Non-ionizing radiation.
1. Ionizing radiation
Ionizing radiations are those which when pass through matter, transfer energy to the
matter rendering it to lose electrons. Ionizing radiation affected atoms of the matter becomes
positively charged particles called ions. Because of these, the molecule containing the positively
charged particles undergoes chemical change. When this change occurs in DNA molecule, the
result is a heritable change-mutation. There are different types of ionizing radiations.
A. Non particulate (Electromagnetic and sparsely ionizing)
 X-rays: They are produced by X-ray machines. They are sparsely ionizing, non
particulate and penetrating. Hard X-rays have wave lengths of 0.1 to 0.01 Å and soft x-
rays have 1 to 10 Å wave lengths.
60
 Gamma rays: They are produced by Co and other radioactive [Link] are
shorter in wave length than x-rays (0.01 Å) and more penetrating than X rays.
B. Particulate

 Alpha particles: They are produced by radioisotopes of heavier elements. They have
two protons and two neutrons. They are positively charged and less penetrating than
neutrons and beta rays; Densely ionizing.
  Beta particles: They are high energy electrons produced by decay of radioactive
32 35
isotopes like P, S, 3H. They are more penetrating than α particles, but less
penetrating than X rays; Sparsely ionizing.
 Fast and thermal neutrons: They are neutral in charge and produced by cyclotron of
atomic reactors by radioactive decay of heavier elements. They are highly penetrating.
(Isotopes are chemically identical substances having the same no. of protons but
different no. of neutrons. Radioisotopes or radioactive isotopes spontaneously
disintegrate Eg. 32P into an element with less no. of neutrons).

2. Non ionizing radiations

Non ionizing radiations are those which when pass through matter transfer energy to the
matter rendering it‟s electrons to move to higher energy levels (higher orbits). In case of
ionizing radiations, electrons are lost by the matter, but in case of non-ionizing radiations,
electrons are raised to higher energy levels (Excitation). The atoms in excited shows increased
activity.
Eg. UV rays: They are produced by mercury vapour lamp. They are lower in energy and
less penetrating. Hence, thin tissues like pollen of plants or eggs of Drosophila are irradiated
using UV rays to induce mutation. Dimer formation and deamination occur due to UV radiation.
II. Chemical mutagens: Oehlkers (1943) found that mixtures of ethylurethane and potassium
chloride induced translocations in Oenothera. Auerbach and Robson reported the mutagenic
action of mustard gas in 1946. Different categories of chemical mutagenic agents are given
below:

1. Alkaylating agents
Eg. Ethyl Methane Sulphonate (EMS), Methyl Methane Sulphonate (MMS), Ethyl
Ethane Sulphonate and Ethylene Imines
2. Acridine dyes
Eg. Acridine orange, Acriflavin, Proflavin
3. Base analogues
Eg. 5 bromo uracil, 5 Chloro uracil, 2 Amino purine
4. Others
Eg. Nitrous Acid, Hydroxylamine and sodium Azide
Among these, EMS, MMS, DES are frequently used for induced mutagenesis.
 Group of mutagen Name of chemical Mode of action

1. Alkylating Agents Ethyl methane Sulphonate AT GC Transitions

Methyl Methane Sulphonate Transitions
Ethyl Ethane Sulphonate GC AT Transitions
Ethylene Imines Transitions
[Link] Analogues 5 Bromo Uracil AT GC Transitions
2 Amino purine AT GC Transitions
[Link] Dyes Acriflavin, Proflavin Deletion, addition and frame
shifts.
[Link] Nitrous Acid AT GC Transitions
Hydroxylamine GC AT Transitions
Sodium Azide Transitions

Mechanisms of chemical mutagenesis

1. Tautomeric shift: If the chemical mutagens change the amino group (-NH2) into imino
group (- NH) in purines, A (amino adenine) = T pairing in DNA is converted to A (imino
adenine) = C pairing. Similarly C=G pairing is converted to C = A pairing. This is called as
tautomeric shift.
In case of pyrimidines, keto group (C=O) is converted to enol group (COH). Because of this A
= T (keto) pairing in converted to G=T (enol) pairing.
2. Substitution: Base analogues substitute for purine and pyrimidines during nucleotide and
DNA synthesis.
3. Deamination: Amino group (NH2) is converted to keto group(C=O). Adenine is converted to
hypoxanthine and cytosine is converted to uracil.

Mutation detection in Drosophila

The task of finding rare mutations in multicellular organisms is difficult compared with that in
microorganisms. In 1928, Hermann J. Muller developed ClB technique for searching lethal
mutation on the X chromosome in Drosophila.

ClB technique

It is useful for scoring of sex linked recessive lethals. He “constructed” an X chromosome

called ClB. The ClB stock of Drosophila has special X chromosome, a large part of which is
inverted. The inversion act as crossover suppressor in the inverted region and designated as C.
Because, in a female fly carrying this special ClB chromosome and a normal normal X, the X
chromosomes do not recombine. The ClB chromosome also bears l, a recessive lethal gene and
the dominant gene B, which determines the dominant bar-eye phenotype. Both these genes
present within the inverted segment. Hence, l and B are inherited together.

ClB/Y males die because of hemizygosity for the lethal allele, but the chromosome can be
maintained in heterozygous ClB/C+l+B+ females. This special ClB system is used for the
detection of sex linked lethal mutations on the X chromosomes in Drosophila males.

He crossed mutagen treated males with ClB females. In F1, half of the females have ClB
chromosome and bar shape. The remaining half without ClB chromosome and normal eye are
rejected. All the surviving F1 males have normal chromosome from ClB females and those
males receiving ClB chromosome die.

Each F1 bar-eyed ClB female is mated to normal male. Progeny from each matings are kept in a
separate culture bottles. Half of the male progeny from F1 ClB female x normal male mating
receive the ClB chromosome die. The remaining half of the males will get their X chromosome
from their mutagen treated grandfather. If there was a new lethal recessive mutation on an X
chromosome in one of the original male gametes, then the F1 female carrying that chromosome
will not produce any viable male progeny. If a normal X chromosome of F1 ClB female had
lethal mutation, there would be no male in its progeny.
The detection of sex linked recessive lethal in this method is based on presence or absence of
male progeny in the cross between F1 bar-eyed ClB female and normal male.
The frequency of recessive lethal mutations in X chromosome of mutagen treated males =
No. of F1 ClB females producing no males in their progeny
---------------------------------------------------------------------------- x100
Total number of F1 ClB females tested

Mutation breeding
The method of crop improvement utilizing the induced mutation is known as mutation breeding.
Procedure:
Applications

i) New genotypes that are not present in germplasm can be created artificially.
ii) Specific qualitative or quantitative characters can be improved in a variety
iii) F1 can be irradiated to increase the variability further to break the linkage groups.
iv) Interspecific hybrids may be irradiated to induce beneficial translocations.
v) For induction of male sterility induced mutagenesis can be used.

Limitations

i. Large populations are to be screened in M2. Each and every single plant to be
observed - laborious.
ii. Desirable mutants linked with other undesirable traits.
iii. Most mutants are recessive. In polyploids, larger population to be studied to find out
recessive mutants.

Achievements – Varieties developed

Black gram- CO 4
Green gram – CO 4
Red gram –CO 3, CO5
Lablab : CO 10
Turmeric : CO1 (From Erode variety)
BSR 1 (X - ray mutant of CO 1)
BSR 2 (X - ray mutant of Erode local)

**********